Cell processing device cell processing system and methods of use thereof

ABSTRACT

A cell processing device for use in performing one or more unit processes in one or more of cell or gene therapy manufacturing, comprising a cell processing platform fluidly coupled to at least one auxiliary container and to at least one primary container, the cell processing platform comprising a body portion comprising at least one fluid inlet fluidly connected to a fluid outlet, and an auxiliary container port fluidly coupled to the at least one fluid inlet of the body portion, wherein the at least one auxiliary container is received in sealing engagement with the auxiliary container port such that the auxiliary container lumen is fluidly connected with the at least one fluid inlet of the body portion, and a primary container is received in sealingly engagement with the primary container port such that the primary container lumen is fluidly connected with the fluid outlet of the body portion.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a national phase entry under 35 U.S.C. § 371 ofInternational Patent Application PCT/GB2020/050009, filed Jan. 3, 2020,designating the United States of America and published as InternationalPatent Publication WO 2020/141327 A1 on Jul. 9, 2020, which claims thebenefit under Article 8 of the Patent Cooperation Treaty to UnitedKingdom Patent Application Serial No. 1900109.8, filed Jan. 4, 2019,United Kingdom Patent Application Serial No. 1900107.2, filed Jan. 4,2019, United Kingdom Patent Application Serial No. 1900108.0, filed Jan.4, 2019, United Kingdom Patent Application Serial No. 1900111.4, filedJan. 4, 2019.

TECHNICAL FIELD

The disclosure relates to a cell processing device for performing one ormore unit operations in cell and/or gene therapy manufacture and methodsof use thereof.

BACKGROUND

Cell and gene therapy manufacturing processes are often complex andinclude manual or semi-automated steps across several devices. Equipmentsystems used in various steps (i.e., unit operations) of cell-basedtherapeutic products (CTP) manufacturing may include devices for cellcollection, cell isolation/selection, cell expansion, cell washing andvolume reduction, cell storage and transportation. The unit operationscan vary immensely based on the manufacturing model (i.e., autologousversus allogenic), cell type, intended purpose, among other factors. Inaddition, cells are “living” entities sensitive to even the simplestmanipulations (such as differences in a cell transferring procedure).The role of cell manufacturing equipment in ensuring scalability andreproducibility is an important factor for cell and gene therapymanufacturing.

In addition, cell-based therapeutic products (CTP) have gainedsignificant momentum thus there is a need for improved cellmanufacturing equipment for various cell manufacturing procedures, forexample, but not limited to stem cell enrichment, generation of chimericantigen receptor (CAR) T cells, and various cell manufacturing processessuch as collection, purification, gene modification,incubation/recovery, washing, infusion into patient and/or freezing.

The culture or processing of cells typically requires the use of adevice to hold the cells, for example, in an appropriate culture mediumwhen culturing the cells. The known devices include shaker flasks,roller bottles, T-flasks and bags. Such bottles or flasks are widelyused but suffer from several drawbacks. Chief among the problems are therequirement for transfer of cells without contamination when passagingor processing subsequently and the sterile addition of supplements andfactors. The existing cell culture devices require re-supply of culturemedium and oxygen for continued cell growth. Gas permeable cell culturedevices are described in U.S. Pat. No. 8,415,144. However, such devicesalso require transfer of medium and/or cells in and out of the devices.

Collapsible cell processing devices for use in medicine are known; see,for example, U.S. Pat. No. 4,867,172 concerning a blood collector, or WO2008/030597 concerning a canister liner for fluid collection. However,such devices are not fabricated or constructed for use in cell and/orgene therapy manufacturing unit operations (i.e., steps).

A key limiting factor in the production of cells or gene therapies foruse in medicine is the absence of compact, automated closed systems forperforming unit operations without contamination. For example, duringcell culture, upstream or subsequent processing of cells, there is arisk of contamination when making additions to the culture vessel, orwhen removing cells or removing liquid samples. The operating systemsare largely manual and hence expensive to operate. Multiple pieces ofequipment are typically required to cover all of the non-cell culturesteps, which involves many transfers, each of which is an opportunityfor operator errors and contamination to occur. Furthermore withincreasing manual operations comes increasing risk of manual errors andtherefore the current labor-intensive processes lack the robustnessrequired for the manufacture of clinical-grade therapeutics.

There is therefore a need for cell processing devices (e.g., multistepcell processors), which permit such processing, which avoids therequirement for constant movement of cells into fresh devices. Forexample, it would be advantageous if scale-up of cells in culture couldbe achieved without transfer of cells into a larger device as the cellpopulation for any given culture increases.

Previous cell manufacturing devices use complex equipment, which islarge and difficult to assemble. The devices use complex networks oftubing, valves and pumps to link elements of the equipment together.

Provided is an improved cell and/or gene therapy processing equipment,which combines the advantages of the cell culture containers of theearlier applications (PCT/GB2016/051451 and PCT/GB2017/053389) (i.e.,avoiding the need for pumps and the requirement for constant passagingof cells into fresh culture devices, holding vessels, tubes etc.) withthe advantages conferred by having individually configurable cell and/orgene therapy processing devices. Together with an improved, closed cellprocessing unit, the improved device and container described hereinpermit a variety of unit processes to be performed within a singledevice or container having a smaller footprint and being less complexthan existing equipment, as will be explained in more detail herein.Moreover, the cell processing containers described herein may maintainan aseptic connection without the prerequisite of a laminar flowcabinet, a glove box, or the like.

The earlier application (PCT/GB2016/051451) describes a cell culturecontainer in which the wall element, being composed of a flexiblematerial, is compressible with respect to its top and base sections. Thecell culture container described therein is compatible with the cellprocessing unit and device described herein.

In a further earlier application (PCT/GB2017/053389) an improved versionof a cell culture container is described, having at least one inlet andfurther comprising one or more auxiliary containers in fluidcommunication with the primary container. The cell culture containerdescribed therein is improved so as to be compatible with the cellprocessing unit and device described herein. Moreover, a connectionbetween the cell culture container described therein and othercomponents is improved, thereby maintaining an aseptic environmentthrough the connection. In the earlier application (PCT/GB2017/053389),a laminar flow cabinet was required in order to ensure an asepticenvironment during cell and/or gene therapy manufacture and/orprocessing. However, this can increase costs and result in a more laborintensive process. Thus, the present application also aims to provide anaseptic connection between components, irrespective of the surroundingenvironment or atmosphere.

BRIEF SUMMARY

It is an object of certain aspects of the disclosure to provide animprovement over the above described techniques and known art;particularly to provide a cell processing unit, a cell processingplatform, a cell processing device and a cell processing container andsystems that facilitate flexible, compact, low cost, multistep cellprocessing while reducing the risk of contamination.

In accordance with the disclosure there is provided a cell processingdevice for use in one or more unit operations in cell and/or genetherapy manufacture and a cell processing system and method inaccordance with the appended claims.

Also described is a cell processing unit for cell and/or gene therapymanufacture and a cell processing system and method.

Also described is a platform cell processing platform for use in one ormore unit operations in cell and/or gene therapy manufacture and a cellprocessing system and method.

Also described is a cell processing container for use one or more unitoperations in cell and/or gene therapy manufacture, a cell processingsystem comprising a cell processing container and a multi-step method ofperforming one or more unit operations in cell and/or gene therapymanufacture.

Cell Processing Device

According to an aspect of the disclosure there is provided a cellprocessing device for use in performing one or more unit operations incell and/or gene therapy manufacture comprising a cell processingplatform according to the disclosure fluidly coupled to at least onecontainer.

The term “cell processing device” is used to define a cell processingplatform having at least one container coupled thereto. The at least onecontainer may be fluidly coupled thereto.

The term “cell processing platform” is used to define a platform, or aninterface, upon which one or more unit operations in cell and/or genetherapy manufacture or processing may be performed. The terms “cellprocessing platform,” “liquid handling platform,” “platform,” “cellprocessing interface” and “interface” can be used synonymously. In someexamples, the cell processing platform serves as an interface betweencomponents, for example, containers, bioreactors or the like, such thatthe user can manipulate the cell processing platform thereby controllingone or more unit operations in cell and/or gene therapy manufacture orprocessing. The cell processing platform may provide a pathway, forexample, a fluid pathway, through conduits, seals, valves, septa or thelike to provide an interface between components, for example,containers, bioreactors or the like.

In some examples, the cell processing platform may be fluidly coupled toat least one container thereby allowing fluid communicationtherebetween. That is, in some examples, the cell process platformallows the introduction or extraction of one or more fluids to or fromthe at least one container.

In certain embodiments, the cell processing platform is fluidly coupledto at least one auxiliary container.

In certain embodiments, the cell processing platform is fluidly coupledto at least one primary container.

Thus in certain embodiments there is provided a cell processing devicefor use in performing one or more unit operations in cell and/or genetherapy manufacture comprising a cell processing platform fluidlycoupled to at least one auxiliary container and being fluidly coupled toat least one primary container.

In certain embodiments the cell processing platform comprises a bodyportion comprising at least one fluid inlet fluidly connected to a fluidoutlet, and an auxiliary container port fluidly coupled to the at leastone fluid inlet of the body portion, wherein the at least one auxiliarycontainer is received in sealing engagement with the auxiliary containerport such that the auxiliary container lumen is fluidly connected withthe at least one fluid inlet of the body portion, and a primarycontainer is received in sealingly engagement with the primary containerport such that the primary container lumen is fluidly connected with thefluid outlet of the body portion.

The primary container may be regarded as a first container. Theauxiliary container may be regarded as a second, or a secondary,container. Any number of containers may be used.

The term “primary container” is used to define that a container isconnected to a first side, or surface, of the cell processing platform.For example, the term “primary container” may be used to define that thecontainer is attached to a lower side, or surface, of the cellprocessing platform. There may be any number of primary containers.

The primary container may be a bellow-based container, for example, abellow-based bioreactor. That is, the container or bioreactor may bebased on a bellows, i.e., a container or bioreactor including a wallelement comprising a series of Z-folds, or a wall element comprising, orforming, a concertina. The bellow-based container or bioreactor mayinclude a base section, a top section arranged substantially in parallelwith the base section and a wall element arranged between the topsection and the base section and defining an internal lumen of thecontainer or bioreactor. The wall element of the container or bioreactorpreferably is compressible with respect to the top and base sections.The wall element of the container or bioreactor may be composed of aflexible material. The wall element may comprise a series of Z-folds.The wall element may comprise, or form, a bellows. The container maytake the form of a concertina.

The term “auxiliary container” or “secondary container” is used todefine that a container is connected to a second side, or surface, ofthe cell processing platform. For example, the term “auxiliarycontainer” or “secondary container” may be used to define that thecontainer is attached to an upper side, or surface, of the cellprocessing platform. There may be any number of auxiliary containers.

The auxiliary container may be a bellow-based container, for example, abellow-based bioreactor. That is, the container or bioreactor may bebased on a bellows, i.e., a container or bioreactor including a wallelement comprising a series of Z-folds, or a wall element comprising, orforming, a concertina. The bellow-based container or bioreactor mayinclude a base section, a top section arranged substantially in parallelwith the base section and a wall element arranged between the topsection and the base section and defining an internal lumen of thecontainer or bioreactor. The wall element of the container or bioreactorpreferably is compressible with respect to the top and base sections.The wall element of the container or bioreactor may be composed of aflexible material. The wall element may comprise a series of Z-folds.The wall element may comprise, or form, a bellows. The container maytake the form of a concertina.

Generally, the term “container,” or a “cell processing container,” isused to define a container, a receptacle, a volume, a bioreactor, or thelike in which one or more unit operations of cell and/or gene therapymanufacture or processing may be completed.

In certain embodiments, the body portion includes one, that is, asingle, fluid inlet, and one, that is, a single, fluid outlet. Incertain embodiments, the body portion includes one or more fluid inlets,and one or more fluid outlets. In certain embodiments, the body portionincludes one, that is a single, fluid inlet, and a plurality of fluidoutlets. In certain embodiments, the body portion includes a pluralityof fluid inlets, and one, that is, a single, fluid outlet. In certainembodiments, the body portions includes a plurality of fluid inlets, anda plurality of fluid outlets.

In certain embodiments, there may be provided one, that is, a single,primary container and one, that is, a single, auxiliary container. Incertain embodiments, there may be provided one, that is, a single,primary container and a plurality of auxiliary containers. In certainembodiments, there may be provided a plurality of primary containers andone, that is, a single, auxiliary container. In certain embodiments,there may be provided a plurality of primary containers and a pluralityof auxiliary containers.

In certain embodiments the at least one auxiliary container isdetachably connected to the auxiliary container port.

In certain embodiments the primary container is detachably connected tothe primary container port.

In certain embodiments, one or more auxiliary containers are indirectlyfluidly coupled to the auxiliary container port. More specifically, oneor more auxiliary containers may be connected to one another in series.Thus, an auxiliary container may be in fluid communication with afurther auxiliary container, wherein the further auxiliary container isnot in direct fluid communication with the auxiliary container port ofthe cell processing platform. Additionally, or alternatively, the cellprocessing device may further comprise one or more further containers,such as a bioreactor, in direct fluid communication with the primarycontainer but not necessarily with the cell processing platform. In thisway, the cell processing device may provide a multistage bioreactoroperable to perform one or more unit processes in a cell and/or genetherapy manufacturing process.

In certain embodiments the auxiliary container port comprises acontainer receiving sleeve connected to the body portion and beingconfigured to surround at least a portion of the auxiliary container,which portion comprises the fluid outlet of the container.

In certain embodiments the container receiving sleeve comprisesinsulation means configured to maintain the contents of an auxiliarycontainer received in the sleeve at a particular temperature. Morespecifically, the insulation means is a thermal sleeve. Accordingly, anauxiliary container port may be configured to maintain the contents ofan auxiliary container at an optimal temperature. For example, theoptimal temperature may be a cell culture temperature (37 degreesCelsius), or room temperature (22 degrees Celsius), or refrigerated(e.g., around 4 degrees Celsius), or below freezing (e.g., around minus4 degrees Celsius or lower, such as minus 20 degrees Celsius, or minus80 degrees).

In certain embodiments, the cell processing device comprises one or moreauxiliary container ports configured to maintain a variety oftemperatures.

In certain embodiments the cell processing platform comprises aplurality of auxiliary container ports and wherein each one of aplurality of auxiliary containers are received in sealing engagementwith one of the plurality of auxiliary container ports such that thelumen of each auxiliary container is fluidly coupled with a fluid inletof the body portion.

In certain embodiments, the auxiliary containers are detachably mountedto the auxiliary container ports.

In certain embodiments each auxiliary container port is coupled to aseparate fluid inlet of the body portion.

In certain embodiments each separate fluid inlet of the body portion isfluidly connected to a fluid outlet of the body portion.

In certain embodiments the at least one fluid inlet and the fluid outletof the body portion are fluidly coupled to one another by a fluidconduit.

In certain embodiments the fluid conduit comprises a valve operable toopen and close the fluid conduit.

In certain embodiments the valve is one of: a pinch valve, a pressuresensitive valve, a clamp valve, a membrane valve, a rupture disc, avenous valve and an aperture valve.

In certain embodiments each auxiliary container port comprises acontainer filling port.

In certain embodiments the container filling port is fluidly connectedto a fluid inlet of the auxiliary container port.

In certain embodiments each container filling port comprises a valveoperatively coupled to the fluid inlet and a fluid outlet of theauxiliary container port and operable to control fluid flow directionthrough the auxiliary container port.

In certain embodiments the container filling port comprises a valveoperable, in an open position, to allow fluid to flow to the fluid inletof the auxiliary container port and not to the fluid outlet of theauxiliary container port and, in a closed position, to close thecontainer filling port and to allow fluid to flow from the fluid inletof the auxiliary container port to the fluid outlet of the auxiliarycontainer port.

In certain embodiments the at least one auxiliary container comprises amating element configured to fluidly connect to a corresponding matingelement on the auxiliary container port.

In certain embodiments the mating element is one of: a sterile connectorend or a LUER-LOK™.

In certain embodiments the primary container port comprises a matingelement configured to fluidly connect to a corresponding mating elementon the primary container.

In certain embodiments the mating element comprises one of: a sterileconnector end or a LUER-LOK™.

In certain embodiments the auxiliary container port comprises aLUER-LOK™ connector at the fluid inlet and/or the fluid outlet of theauxiliary container port, each LUER-LOK™ connector configured to engagewith a further LUER-LOK™ connector on a container and/or on the bodyportion, respectively. More specifically, a male LUER-LOK™ connector isconfigured to engage with a female LUER-LOK™ connector.

In certain embodiments the fluid outlet of the body portion comprises aLUER-LOK™ connector configured to engage with a further LUER-LOK™connector on a primary container attachable to the body portion.

In certain embodiments the auxiliary container port comprises a sterileconnector end at the fluid inlet and/or the fluid outlet of theauxiliary container port, each sterile connector end configured toengage with a further sterile connector end on a container and/or on thebody portion, respectively.

In certain embodiments the fluid outlet of the body portion comprises asterile connector end configured to engage with a further sterileconnector end on the primary container attachable to the body portion.

In certain embodiments the cell processing device comprises at least onepositional tracking device operable to indicate a set location on thecell processing platform. In this way, the position of the platform maybe tracked, for example, when the cell processing device is mounted intoa cell processing unit according to the disclosure.

In certain embodiments the at least one positional tracking device is amechanical device.

In certain embodiments, the at least one positional tracking devicecomprises a cog. In such embodiments, the mounting plate of the cellprocessing unit may comprise a further cog operable to engage theprojections of the cog on the cell processing platform. In this way, thecell processing device will need to be physically inserted into themounting plate of the cell processing unit in the correct orientation.This, in turn, ensures the operator knows the position of the device andthus containers mounted to the platform in the cell processing unit.

In certain embodiments the positional tracking device is an encoder.More specifically, the positional tracking device is one or more of: amagnet, an RFID sensor, a light sensor or the like.

In certain embodiments the cell processing device comprises a pluralityof positional tracking devices.

In certain embodiments the at least one positional tracking device islocated on the cell processing platform relative to the auxiliarycontainer port such that the location of the positional tracking deviceis related to the position of the auxiliary container port.

In certain embodiments the at least one positional tracking device islocated on the body portion of the cell processing platform relative tothe auxiliary container port.

In certain embodiments the system comprises a plurality of positionaltracking devices each located on the body portion of the cell processingplatform relative to an auxiliary container port.

In certain embodiments the cell processing device comprises a samplingport in the body portion of the cell processing platform. Alternatively,the sampling port may be located in the base section of the primarycontainer.

In certain embodiments the cell processing device comprises a gastransfer port in the body portion of the cell processing platform.Alternatively, the gas transfer port may be located in the wall of theprimary container.

In certain embodiments the auxiliary container port is configured toreceive a container having a base section, a top section arrangedsubstantially in parallel with the base section and a wall elementarranged between the top section and the base section and defining aninternal lumen of the container, in which the wall element of thecontainer preferably is compressible with respect to the top and basesection and the wall element of the container is composed of a flexiblematerial.

In certain embodiments the primary container port is configured toreceive a primary container having a base section, a top sectionarranged substantially in parallel with the base section and a wallelement arranged between the top section and the base section anddefining an internal lumen of the container, in which the wall elementof the container preferably is compressible with respect to the top andbase section and the wall element of the container is composed of aflexible material.

In certain embodiments the primary container further comprises anattachment flange mounted to the top section of the primary containerand being configured to sealingly engage and detachably mount to theprimary container port.

In certain embodiments the at least one auxiliary container iscompressible. In this way, the container configuration is based on aconcertina (which can act as a pump) therefore there is no need forseparate pumps and complex sets of tubing/pipes to transfer the contentsof a container to another container in the system. In turn thisconfiguration reduces the space needed for a cell and/or gene therapymanufacturing process.

In certain embodiments, the container is a container described in theearlier patent application PCT/GB2016/051451.

In certain embodiments, the container is a container described in theearlier patent application PCT/GB2017/053389.

Alternatively the container may comprise a syringe arrangement allowingit to be re-filled or emptied.

In certain embodiments the at least one auxiliary container is asyringe. In such a syringe arrangement, the container has an arrangementanalogous to a syringe having an element that is moveable to eitherexpel fluid from the container or draw it back in.

In certain embodiments, the container may comprise any shaped containerwith a moving seal allowing variable volume operations.

In certain embodiments the at least one auxiliary container is a bagretained in a frame and moveable with respect to the frame. Morespecifically, the top section, the base section and wall element of theat least one auxiliary container may form a bag, which can be heldwithin an external adjustable frame, or in which the bag comprises aninternal adjustable frame within the material of the bag. Accordingly,one or more of the auxiliary containers in fluid communication with acell processing platform of the disclosure may form a bag, which can beheld within an external adjustable frame, or in which the bag comprisesan internal adjustable frame within the material of the bag. Such a bagmay be configured to act, for example, as an intravenous drip bag. Itwill therefore be understood the product(s) of any reaction(s) carriedout in a primary container or further container of the cell processingdevice may be collected into the bag, which can then be removed andtransferred to an intravenous drip. Alternatively, the product(s) of anyreaction(s) can be directly delivered to a patient from the lumen of thecontainer,

In certain embodiments the cell processing device comprises one or moreauxiliary containers connected to an auxiliary container port of thecell processing platform. More specifically, the one or more auxiliarycontainers are detachably connected to an auxiliary container port ofthe cell processing platform.

In certain embodiments one or more of the auxiliary containers areconnected to a respective auxiliary container port with a sterileconnector.

In certain embodiments one or more of the auxiliary containers areconnected to a respective auxiliary container port with a LUER-LOK™style connector.

In certain embodiments the at least one auxiliary container is locatedon the top of the cell processing platform.

In certain embodiments the primary container is located on the bottom ofthe cell processing platform.

According to a further aspect, the disclosure provides a multi-stepmethod of performing one or more unit operations in cell and/or genetherapy manufacture using a cell processing device according to thedisclosure.

In certain embodiments the method comprises introducing a cellpopulation, for example, a cell population of interest, into the primarycontainer and sequentially adding one or more reagents from one or moreauxiliary containers into the primary container via the cell processingplatform in order to effect growth, culturing and/or modification of thecells, for example, in order to effect a desired growth, culturingand/or modification of the cells.

In certain embodiments the auxiliary container is one of: a reagentcontainer, a cell culture container, a waste container, an emptycontainer or a bioreactor.

In certain embodiments the primary container is one of: a cell culturecontainer or a bioreactor, a reagent container, a waste container, afilter, an electroporator, a purifier, a waste container, a filter, anelectroporator, a purifier, holding container, apheresis/leukopheresis,differentiation chamber, chromatography column, settling chamber, sieve,shaking/mixer, a centrifuge and a magnetic bead separator or the like.

A container of the disclosure may be of circular, square, rectangular,elliptical, or triangular cross section. Alternatively, a container ofthe disclosure may comprise a number of different sections or regions ofa variety of cross sections, such as, for example, a series of circularcross sections with variable (increasing and /or decreasing) diameters.

Advanced blow molding techniques can be used to deposit a second (oreven third), external, coating or layer of plastic impermeable to oxygenonto the wall, top and base of the auxiliary container. In this way,shelf life of the container in storage can be extended.

According to a yet further aspect the disclosure provides a cellprocessing system comprising a cell processing device according to anaspect of the disclosure and a cell processing unit according to thedisclosure.

Cell Processing Platform

According to an aspect of the disclosure there is provided a cellprocessing platform for use in performing one or more unit operations incell and/or gene therapy manufacture, the platform comprising a bodyportion comprising at least one fluid inlet fluidly connected to a fluidoutlet, and an auxiliary container port fluidly coupled to the at leastone fluid inlet of the body portion, wherein the auxiliary containerport is configured and arranged to receive and sealingly engage with anauxiliary container and to fluidly connect the auxiliary container lumenwith the at least one fluid inlet of the body portion, and a primarycontainer port configured and arranged to sealingly engage with aprimary container and to fluidly connect the primary container lumenwith the fluid outlet of the body portion.

The term “cell processing platform” is used to define a platform, or aninterface, upon which one or more unit operations in cell and/or genetherapy manufacture or processing may be performed. The terms “cellprocessing platform,” “liquid handling platform,” “platform,” “cellprocessing interface” and “interface” can be used synonymously. In someexamples, the cell processing platform serves as an interface betweencomponents, for example, containers, bioreactors or the like, such thatthe user can manipulate the cell processing platform thereby controllingone or more unit operations in cell and/or gene therapy manufacture orprocessing. The cell processing platform may provide a fluid pathway,through conduits, seals, valves, septa or the like to provide aninterface between components, for example, containers, bioreactors orthe like. The cell processing platform may provide an aseptic fluidpathway through conduits, seals, valves, septa or the like to provide aninterface between components, for example, containers, bioreactors orthe like.

The primary container may be regarded as a first container. Theauxiliary container may be regarded as a second, or a secondary,container. Any number of containers may be used.

The term “primary container” is used to define that a container isconnected to a first side, or surface, of the cell processing platform.For example, the term “primary container” may be used to define that thecontainer is attached to a lower side, or surface, of the cellprocessing platform. There may be any number of primary containers.

The primary container may be a bellow-based container, for example, abellow-based bioreactor. That is, the container or bioreactor may bebased on a bellows, i.e., a container or bioreactor including a wallelement comprising a series of Z-folds, or a wall element comprising, orforming, a concertina. The bellow-based container or bioreactor mayinclude a base section, a top section arranged substantially in parallelwith the base section and a wall element arranged between the topsection and the base section and defining an internal lumen of thecontainer or bioreactor. The wall element of the container or bioreactorpreferably is compressible with respect to the top and base sections.The wall element of the container or bioreactor may be composed of aflexible material. The wall element may comprise a series of Z-folds.The wall element may comprise, or form, a bellows. The container maytake the form of a concertina.

The term “auxiliary container” or “secondary container” is used todefine that a container is connected to a second side, or surface, ofthe cell processing platform. For example, the term “auxiliarycontainer” or “secondary container” may be used to define that thecontainer is attached to an upper side, or surface, of the cellprocessing platform. There may be any number of auxiliary containers.

The auxiliary container may be a bellow-based container, for example, abellow-based bioreactor. That is, the container or bioreactor may bebased on a bellows, i.e., a container or bioreactor including a wallelement comprising a series of Z-folds, or a wall element comprising, orforming, a concertina. The bellow-based container or bioreactor mayinclude a base section, a top section arranged substantially in parallelwith the base section and a wall element arranged between the topsection and the base section and defining an internal lumen of thecontainer or bioreactor. The wall element of the container or bioreactorpreferably is compressible with respect to the top and base sections.The wall element of the container or bioreactor may be composed of aflexible material. The wall element may comprise a series of Z-folds.The wall element may comprise, or form, a bellows. The container maytake the form of a concertina.

Generally, the term “container,” or a “cell processing container,” isused to define a container, a receptacle, a volume, a bioreactor, or thelike in which one or more unit operations of cell and/or gene therapymanufacture or processing may be completed.

In certain embodiments, the body portion includes one, that is, a singlefluid inlet, and one, that is, a single, fluid outlet. In certainembodiments, the body portion includes one or more fluid inlets, and oneor more fluid outlets. In certain embodiments, the body portion includesone, that is a single, fluid inlet, and a plurality of fluid outlets. Incertain embodiments, the body portion includes a plurality of fluidinlets, and one, that is, a single, fluid outlet. In certainembodiments, the body portions includes a plurality of fluid inlets, anda plurality of fluid outlets.

In certain embodiments the auxiliary container port comprises a sealablefluid inlet and/or a sealable fluid outlet.

In certain embodiments, the auxiliary container port is configured forsealing engagement with the fluid outlet of an auxiliary container.

In certain embodiments, the primary container port is configured forsealing engagement with the fluid inlet of a primary container.

In certain embodiments the auxiliary container port comprises acontainer receiving sleeve connected to the body portion and beingconfigured to surround at least a portion of the auxiliary container,which portion comprises the fluid outlet of the container.

In certain embodiments the container receiving sleeve comprisesinsulation means configured to maintain the contents of an auxiliarycontainer received in the sleeve at a particular temperature. Morespecifically, the insulation means is a thermal sleeve. Accordingly, theauxiliary container receiving port may be configured to maintain thecontents of a container received within the port at an optimaltemperature. For example, the optimal temperature may be cell culturetemperature (37 degrees Celsius), or room temperature (22 degreesCelsius), or refrigerated (e.g., around 4 degrees Celsius), or belowfreezing (e.g., around minus 4 degrees Celsius or lower, such as minus20 degrees Celsius, or minus 80 degrees).

In certain embodiments the cell processing platform may have one or moreauxiliary container ports configured to maintain a variety oftemperatures.

In certain embodiments the auxiliary container port comprises a matingelement configured to fluidly connect to a corresponding mating elementon an auxiliary container.

In certain embodiments the mating element is one of: a sterile connectorend or a LUER-LOK™. When the mating element of the auxiliary containerport is a LUER-LOK™ the port may have a male LUER-LOK™ connector, whichwill engage and couple with a corresponding female LUER-LOK™ connectoron the container or vice versa.

In certain embodiments the primary container port comprises a matingelement configured to fluidly connect to a corresponding mating elementon a primary container.

In certain embodiments the mating element comprises one of: a sterileconnector end or a LUER-LOK™ When the mating element of the primarycontainer port is a LUER-LOK™, the port may have a male LUER-LOK™connector, which will engage and couple with a corresponding femaleLUER-LOK™ connector on the container or vice versa.

In certain embodiments the auxiliary container port comprises aLUER-LOK™ connector at the fluid inlet and/or the fluid outlet of theauxiliary container port, each LUER-LOK™ connector configured to engagewith a further LUER-LOK™ connector on a container and/or on the bodyportion, respectively. More specifically, a male LUER-LOK™ connector isconfigured to engage with a female LUER-LOK™ connector.

In certain embodiments the fluid outlet of the body portion comprises aLUER-LOK™ connector configured to engage with a further LUER-LOK™connector on a primary container attachable to the body portion.

In certain embodiments the auxiliary container port comprises a sterileconnector end at the fluid inlet and/or the fluid outlet of theauxiliary container port, each sterile connector end configured toengage with a further sterile connector end on a container and/or on thebody portion, respectively.

In certain embodiments the fluid outlet of the body portion comprises asterile connector end configured to engage with a further sterileconnector end on a primary container attachable to the body portion.

In certain embodiments the body portion is substantially hollow.

In certain embodiments the at least one fluid inlet and the fluid outletof the body portion are fluidly coupled to one another by a fluidconduit.

In certain embodiments the fluid conduit comprises a valve operable toopen and close the fluid conduit.

In certain embodiments the valve is one of: a pinch valve, apressure-sensitive valve, a clamp valve, a membrane valve, a rupturedisc, a venous valve and an aperture valve.

In certain embodiments the auxiliary container port comprises acontainer filling port.

In certain embodiments the container filling port is fluidly connectedto a fluid inlet of the auxiliary container port.

In certain embodiments the container filling port comprises a valveoperatively coupled to the fluid inlet and a fluid outlet of theauxiliary container port and operable to control fluid flow directionthrough the auxiliary container port.

In certain embodiments the container filling port comprises a valveoperable, in an open position, to allow fluid to flow to the fluid inletof the auxiliary container port and not to the fluid outlet of theauxiliary container port and, in a closed position, to close thecontainer filling port and to allow fluid to flow from the fluid inletof the auxiliary container port to the fluid outlet of the auxiliarycontainer port.

In certain embodiments the platform comprises a plurality of auxiliarycontainer ports each fluidly connected to a fluid inlet of the bodyportion. In this way, each of the plurality of auxiliary container portsis configured and arranged to receive and sealingly engage with anauxiliary container and to fluidly connect the container lumen with afluid inlet of the body portion.

In certain embodiments each auxiliary container port is coupled to aseparate fluid inlet of the body portion.

In certain embodiments each separate fluid inlet of the body portion isfluidly connected to a fluid outlet of the body portion.

In certain embodiments the platform comprises at least one positionaltracking device operable to indicate a set location on the platform. Inthis way, the position of the platform may be tracked, for example, whenthe platform is mounted into a cell processing unit according to thedisclosure.

In certain embodiments the at least one positional tracking device is amechanical device.

In certain embodiments, the at least one positional tracking devicecomprises a cog. In such embodiments, the mounting plate of the cellprocessing unit may comprise a further cog operable to engage theprojections of the cog on the cell processing platform. In this way, thecell processing platform will need to be physically inserted into themounting plate of the cell processing unit in the correct orientation.This, in turn, ensures the operator knows the position of the platformand thus containers mounted to the platform in the cell processing unit.

In certain embodiments the positional tracking device is an encoder.More specifically the positional tracking device is one or more of: amagnet, an RFID sensor, a light sensor or the like.

In certain embodiments the platform comprises a plurality of positionaltracking devices.

In certain embodiments the at least one positional tracking device islocated relative to the (or each) auxiliary container port such that thelocation of the positional tracking device is related to the position ofthe auxiliary container port.

In certain embodiments the at least one positional tracking device islocated on the body portion relative to the auxiliary container port.

In certain embodiments the platform comprises a sampling port in thebody portion.

In certain embodiments the platform comprises a gas transfer port in thebody portion.

In certain embodiments the auxiliary container port is configured toreceive a container having a base section, a top section arrangedsubstantially in parallel with the base section and a wall elementarranged between the top section and the base section and defining aninternal lumen of the container, in which the wall element of thecontainer preferably is compressible with respect to the top and basesection and the wall element of the container is composed of a flexiblematerial.

In certain embodiments, the auxiliary container port is configured toreceive a container described in International Patent Application NumberPCT/GB2016/051451.

In certain embodiments, the auxiliary container is detachably mounted tothe auxiliary container port.

In certain embodiments the primary container port is configured toreceive a primary container having a base section, a top sectionarranged substantially in parallel with the base section and a wallelement arranged between the top section and the base section anddefining an internal lumen of the container, in which the wall elementof the container preferably is compressible with respect to the top andbase section and the wall element of the container is composed of aflexible material.

In certain embodiments, the auxiliary container port is configured toreceive a primary container described in International PatentApplication Number PCT/GB2016/051451 or PCT/GB2017/053389.

In certain embodiments the primary container further comprises anattachment flange mounted to the top section of the primary containerand being configured to sealingly engage and mount to the primarycontainer port.

In certain embodiments, the primary container is detachably mounted tothe primary container port.

Cell Processing Container

A critical step, and risk, in performing unit operations in cell and/orgene therapy manufacture, is the sterile connection of the components ofthe equipment to form a usable cell processing device or the like.

At least this object and advantages that will be apparent from thedescription have been achieved by a cell processing container for use inone or more unit operations in cell and/or gene therapy manufacture, thecontainer having a base section, a top section arranged substantially inparallel with the base section and a wall element arranged between thetop section and the base section and defining an internal lumen of thecontainer, in which the wall element of the cell culture containerpreferably is compressible with respect to the top and base section andthe wall element of the cell culture container is composed of a flexiblematerial, wherein the cell processing container comprises at least onesterile connector end configured to operatively couple with a furthersterile connector end so as to form a sterile connector between the cellprocessing container and a further component to which the cellprocessing container is to be fluidly connected.

A sterile connector when referred to herein shall at least include asterile connecting device configured to produce a sterile connection orsterile welds between two elements, for example, two containers or twopieces of compatible tubing. This procedure permits sterile connectionof a variety of containers and tubes of varying diameters by maintaininga closed system as the two portions of the sterile connecting device aremated with one another. In this way, a sterile fluid pathway ismaintained between two elements, for example, containers, tubes or thelike. Each tube/container may have a sterile connector end embeddedtherein and may have a removable membrane (e.g., paper) or valve barrierfor mating to another connector end embedded in a furthertube/container. Sterile connectors are designed to connect oneprocessing stream to another, such as a container to a sampling line,media to a product vessel, or a filtration assembly to a filling line.They become beneficial when no biocontainment hood is available to makean aseptic connection as, owing to the aseptic pathway created, asterile connection can be achieved irrespective of the environment ofsurroundings in which the connection is made.

The line at the junction of the connection cannot be disconnectedwithout force because of safety mechanisms in place to prevent this.Disconnection between connected sterile connector ends may, for example,require a disconnection device, tube sealer, or tube crimper.

The term “fluidly connected” is used to refer to a connection betweencomponents to allow passageway of a fluid. The term “fluid” is used torefer to gases and liquids, in addition to solutions, suspensions,pastes and gels. Moreover, fluid may also refer to granularparticulates, or solids, such as powders. Such particulates, solids orpowders may or may not be suspended within a liquid, as a solution, orthe like.

In some examples, the wall element of the cell processing containerpreferably, that is, optionally, is compressible with respect to the topand base section. That is, the top section may be compressed withrespect to the base section, or the base section may be compressed withrespect to the top section, or the top section and the base section maybe compressed with respect to the base section and the top section,respectively.

In other examples, the wall element of the cell processing container isnot compressible with respect to the top and base sections. In someexamples, the wall element of the cell processing container may beflexible, such that a compression, or squeezing, of the wall elementinwardly toward a central longitudinal axis may be achieved.

Generally, the term “container,” or “cell processing container,” is usedto define a container, a receptacle, a volume, a bioreactor, or the likein which one or more unit operations of cell and/or gene therapymanufacture or processing may completed.

In certain embodiments the at least one sterile connector end is agenderless sterile connector end configured to operatively couple with afurther genderless sterile connector end.

That is, the at least one sterile connector end may be genderless in thesense that it includes neither a male portion nor a female portion. Insome examples, the genderless sterile connector end may include one ormore portions that cooperate with a portion of a further genderlesssterile connector end.

In certain embodiments the at least one sterile connector end is a malesterile connector end configured to operatively couple with a femalesterile connector end.

In certain embodiments the at least one sterile connector end is afemale sterile connector end configured to operatively couple with amale sterile connector end.

In certain embodiments the cell processing container comprises aplurality of sterile connector ends each configured to operativelycouple with a separate further sterile connector end to form a pluralityof sterile connectors between the cell processing container and at leastone further component to which the cell processing container is to befluidly connected.

In certain embodiments the at least one further component is one of: afurther cell processing container, a cell processing platform accordingto the disclosure, a tube or the like.

In certain embodiments the sterile connector ends are embedded in thecell processing container.

In some examples, the sterile connector ends may form part of the cellprocessing container. In some examples, the sterile connector ends mayform an integral part, or may be integrally formed within, or as partof, the cell processing container.

In certain embodiments the sterile connector end is operatively coupledto a pinch valve embedded in the cell processing container.

In certain embodiments the cell processing container has a circular,square, rectangular, elliptical, or triangular cross section.

In certain embodiments, when the cell processing container has acircular shape, the sterile connector end(s) is/are connected to the topand/or base section of the cell processing container in an essentiallycircular pattern.

According to an aspect of the disclosure there is provided a cellprocessing system, comprising a cell processing container as describedabove, further comprising one or more auxiliary containers detachablyconnected to the cell processing container.

In certain embodiments one or more of the auxiliary containers comprisesthe further sterile connector end and is connected to the cellprocessing container via the further sterile connector end.

In certain embodiments one or more of the auxiliary containers islocated at or near the top section of the cell processing container.

In certain embodiments, one or more of the auxiliary containers islocated on the top section of the cell processing container.

In certain embodiments, one or more of the auxiliary containers islocated on the top section of the cell culture container.

In certain embodiments one or more of the auxiliary containers islocated at or near the base section of the cell processing container.

In certain embodiments one or more auxiliary containers is located onthe top section of the cell processing container.

In certain embodiments one or more of the auxiliary containers islocated on the base section of the cell culture container.

In certain embodiments one or more containers may be connected inseries. For example, the cell processing system of the disclosure maycomprise an auxiliary container, which is in fluid communication with afurther auxiliary container, wherein the further auxiliary container isnot is direct fluid communication with the cell processing container ofthe system.

In certain embodiments each container in a series of containerscomprises a sterile connector end in a top and in a base section. Inthis way, it is possible to undertake one or more processing steps in anauxiliary container before making a sterile connection via the sterileconnector ends in connected containers in order to undertake one or morefurther processing steps in the combined containers. In certainembodiments, the one or more processing steps and the one or morefurther processing steps may involve different cell processing units.

In certain embodiments the one or more auxiliary containers have a basesection, a top section arranged substantially in parallel with the basesection and a wall element arranged between the top section and the basesection and defining an internal lumen of the container, in which thewall element of the auxiliary container preferably is compressible withrespect to the top and base section and the wall element of theauxiliary container is composed of a flexible material.

Advanced blow molding techniques can be used to deposit a second (oreven third), external, coating or layer of plastic impermeable to oxygenonto the wall, top and base of the auxiliary container. In this way,shelf life of the container in storage can be extended.

According to a yet further aspect of the disclosure there is provided acell processing system operable to perform one or more unit operationsin cell and/or gene therapy manufacture. The cell processing systemcomprises, a cell processing unit according to an aspect of thedisclosure, a cell processing device according to an aspect of thedisclosure comprising a cell processing platform according to an aspectof the disclosure.

In certain embodiments, the cell processing system comprises at leastone cell processing container according to an aspect of the disclosure.

According to a yet further aspect of the disclosure there is provided amulti-step method of one or more unit operations in cell and/or genetherapy manufacture using a cell culture system according to thedisclosure.

In certain embodiments the method comprises introducing a cellpopulation, for example, a cell population of interest, into the cellprocessing container and sequentially adding one or more reagents fromone or more auxiliary containers into the cell processing container inorder to effect one or more unit operations in cell and/or gene therapymanufacture, for example, desired one or more unit operations in celland/or gene therapy manufacture.

Cell Processing Unit

According to an aspect of the disclosure there is provided a cellprocessing unit for cell and gene therapy manufacture comprising ahousing defining an enclosure into which a cell processing platform canbe mounted, a platform mounting bracket within the housing andconfigured and arranged to receive and retain a cell processingplatform, a drive apparatus configured and arranged to operativelyengage and act upon the cell processing platform so as to move same withrespect to the platform mounting bracket, and an actuator configured andarranged to exert a force on a container mounted into the cellprocessing platform so as to expel a contents from the container.

The term “cell processing unit” is used to define a unit in which one ormore unit operations in cell and/or gene therapy manufacture orprocessing may be performed. The cell processing unit may serve as ahousing for components used in such manufacture and processing. The cellprocessing unit may take any suitable shape or size. The cell processingunit may take the form of an apparatus or the like. That is, the terms“cell processing unit,” “cell processing apparatus” and “an apparatusfor cell and/or gene therapy manufacture or processing” may be usedinterchangeably.

The term “enclosure” is used to define an area or space in which anothercomponent can be received, housed or enclosed, either partially orfully. The enclosure may take the form of a chamber, a receptacle, avolume of space or the like.

The term “cell processing platform” is used to define a platform, or aninterface, upon which one or more unit operations in cell and/or genetherapy manufacture or processing may be performed. The terms “cellprocessing platform,” “liquid handling platform,” “platform,” “cellprocessing interface” and “interface” can be used synonymously. In someexamples, the cell processing platform serves as an interface betweencomponents, for example, containers, bioreactors or the like, such thatthe user can manipulate the cell processing platform thereby controllingone or more unit operations in cell and/or gene therapy manufacture orprocessing. The cell processing platform may provide a fluid pathway,through conduits, seals, valves, septa or the like to provide aninterface between components, for example, containers, bioreactors orthe like.

The term “platform mounting bracket” is used to define a mountingbracket for a cell processing platform as described herein. The platformmounting bracket may take the form of one or more components configuredand arranged such that a cell processing platform may be mountedthereto.

The term “actuator” is used to define an operable mechanism that maycause actuation, or operation, of one or more components of the cellprocessing unit or cell processing platform. In some examples, theactuator may cause actuation, or operation, of one or more containers.In some examples, the actuator may cause actuation, or operation orcompression, of one or more compressible containers. In some examples,the actuator may cause actuation, or operation, of one or more valves.

In certain embodiments the housing is accessible through a door in awall of the housing. More specifically, the door may be hingedlyconnected to the wall of the housing. Yet more specifically, the door ispositioned in a front wall of the housing. In this way, front loading ofthe cell processing unit is possible.

In certain embodiments, the housing has a rectangular or squarefootprint.

In certain embodiments the platform mounting bracket comprises amounting plate. More specifically, the mounting plate is configured toreceive a portion of a cell processing platform. In this way, a cellprocessing platform is retained on the mounting plate when in use.

In certain embodiments the platform mounting bracket comprises aretaining flange spaced apart from the mounting plate in order that acell processing platform can be received and retained in position in thehousing between the mounting plate and the retaining flange. Morespecifically, the retaining flange and the mounting plate togetherprovide a recess (slot) into which a portion of a cell processingplatform can be located and retained.

In certain embodiments the mounting plate is substantially C-shaped.Thus, a cell processing platform can be moved into location on themounting plate from a sideways (i.e., front) loading position.

In certain embodiments the mounting plate is mounted to the housing.

In certain embodiments, the mounting plate is adjustable. Morespecifically, the distance between the base of the housing and themounting plate is adjustable. In this way, different cell processingdevices can be located in the housing.

In certain embodiments the mounting plate is positioned within thehousing to allow a cell processing device to be supported by the platewithout contacting the walls, top or base of the housing. In this way,the mounting plate suspends a cell processing device in the housing. Thecell processing device is therefore able to rotate in the housing.

In certain embodiments the drive apparatus is a rotational driveapparatus configured and arranged to operatively engage and act upon acell processing platform so as to rotate same with respect to theplatform mounting bracket. Thus, the cell processing platform, oncepositioned in the cell processing unit and engaged with the rotationaldrive apparatus, can be indexed in its position relative to the platformmounting bracket by operation of the rotational drive apparatus. Thus,in certain embodiments the cell processing unit is operable to move acell processing platform within it in an automatic process.

In certain embodiments the rotational drive apparatus comprises a drivewheel, which is mounted on the platform mounting bracket and isconfigured to engage a surface of a cell processing platform and toimpart rotational movement on it.

In certain embodiments the rotational drive apparatus comprises a sprungwheel biased toward the drive wheel and spaced apart from it and mountedon the platform mounting bracket.

In certain embodiments the rotational drive apparatus comprises a hingedwheel biased toward the drive wheel and spaced apart from it and mountedon the platform mounting bracket.

The term “hinged wheel” is used to define a wheel hingedly mounted, thatis, mounted upon a hinge, such that it may be moveable between at leasta first configuration and a second configuration. The wheel may behingedly mounted in any suitable way, and may be moveable between anynumber of appropriate configurations.

In certain embodiments the hinged wheel is moveable into an openposition in which a cell processing platform can be inserted into andengaged with the platform mounting bracket and a closed position inwhich the hinged wheel is engaged with a surface of the cell processingplatform in order to retain same in the cell processing platformmounting bracket.

In certain embodiments the hinged wheel is moveable manually.

In certain embodiments the hinged wheel is moveable automatically. Morespecifically, the hinged wheel may be operatively coupled to an actuatoroperable to move the hinged wheel between the open position and theclosed position.

In certain embodiments the hinged wheel is mounted to the door of thehousing.

In certain embodiments the door of the housing comprises a platformengaging means. More specifically, the platform engaging means is oneof: a flange, a protrusion or a lug located on the inside of the door(facing the inside of the housing) and being operable to engage with thesurface of a cell processing platform when the door is closed. In thisway, the platform engaging means is operable to retain the cellprocessing platform in the mounting bracket. The platform engaging meansmay also be operable to maintain the cell processing platform inengagement with the drive wheel of the rotational drive apparatus.

In certain embodiments the drive apparatus comprises a three-pointcontact arrangement. In this way, a cell processing platform in the cellprocessing unit is retained in the drive mechanism around its fullcircumference.

In certain embodiments, the three elements of the drive apparatus (e.g.,the drive wheel, the sprung wheel and the hinged wheel) are equallyspaced from one another within the housing. Such an arrangementfacilitates the rotational movement of the cell processing platform withthe least number of drive wheels.

In certain embodiments the actuator is a linear actuator.

The term “linear actuator” is used to define an actuator that moves in alinear manner, that is, along an axis. In some examples, the linearactuator may be operable along a longitudinal axis.

In certain embodiments, the actuator comprises a lever, a plunger or aseries of levers, plungers or bellows configured to compress a containermounted into the cell processing platform. In certain embodiments theactuator is configured to compress the primary container and/or the oneor more auxiliary containers mounted to the cell processing platform andlocated in the housing.

In certain embodiments the linear actuator comprises a plungeroperatively coupled to a drive motor, wherein the plunger is configuredto engage a container in the cell processing platform and to exert acompression force on the container.

In certain embodiments, the cell processing unit comprises a pluralityof actuators. In certain embodiments the apparatus comprises a primaryactuator configured and arranged to exert a force on a primary containermounted to the cell processing platform so as to expel a contents (e.g.,a fluid, cells or the like) from the container.

In certain embodiments the primary actuator is a linear actuator.

In certain embodiments, the linear primary actuator comprises a lever, aplunger or a series of levers, plungers or bellows configured tocompress the primary container.

In certain embodiments the primary actuator comprises a plungeroperatively coupled to a drive motor, wherein the plunger is configuredto engage a primary container mounted to the cell processing platformand to exert a compression force on the primary container.

It will be understood that any actuator should preferably be capable notmerely of compressing or collapsing a container mounted to the cellprocessing platform but also of re-opening it where this is required. Inthis way, the contents of the container can be agitated by repeatedcompression/extension of the container.

In certain embodiments the apparatus comprises a valve actuator operableto act upon a valve in the cell processing platform so as to open andclose same as force is applied to the container. In certain embodimentsthe valve is a pinch valve.

In certain embodiments the valve actuator is a linear actuator.

In certain embodiments the valve actuator comprises a solenoid valve.

In certain embodiments the apparatus comprises a location detectingsensor operable to detect the position of the cell processing platformrelative to the platform mounting bracket.

In certain embodiments the location detecting sensor is operable todetect the rotational position of the cell processing platform relativeto the platform mounting bracket. In this way, the location of containerports, and therefore the containers mounted in the cell processingplatform, are detectable as the cell processing platform moves relativeto the housing.

In certain embodiments the location detecting sensor comprises one ormore of: a Hall Effect sensor, an RFID sensor, a light sensor or a cogoperable to engage a further cog.

In certain embodiments the apparatus comprises a home location detectingsensor operable to detect a home position of the cell processingplatform relative to the platform mounting bracket.

The term “home position” is used to define a first, default or originalposition of configuration of the cell processing platform. The homeposition may be referred to as so with respect to a predeterminedposition in relation to the platform mounting bracket.

In certain embodiments the home location detecting sensor is operable todetect a single rotational position of the cell processing platformrelative to the platform mounting bracket.

The term “a single rotational position” is used to define a positionwithin the path of rotation of the cell processing platform. The singlerotational position may be referred to as so with respect to a positionon the platform mounting bracket, for example, a predetermined position.

In certain embodiments the home location detecting sensor comprises oneor more of: a Hall Effect sensor, an RFID sensor, a light sensor or acog operable to engage a further cog.

In certain embodiments the voltage detected by the Hall Effect sensor isgreater at the home position of the cell processing platform relative tothe platform mounting bracket than at any other position during therotation of the cell processing platform relative to the platformmounting bracket.

In certain embodiments a container is mounted to the cell processingplatform to form a cell processing device as described herein. Morespecifically, the container is compressible. In this way, the containerconfiguration is based on a concertina (which can act as a pump)therefore there is no need for separate pumps and complex sets oftubing/pipes to transfer the contents of a container to anothercontainer in the system. In turn this configuration reduces the spaceneeded for a cell and/or gene therapy manufacturing process.

In certain embodiments, the container is a container described in theearlier patent application PCT/GB2016/051451.

In certain embodiments, the container is a container described in theearlier patent application PCT/GB2017/053389.

In certain embodiments the container comprises a base section, a topsection arranged substantially in parallel with the base section and awall element arranged between the top section and the base section anddefining an internal lumen of the container, in which the wall elementof the container preferably is compressible with respect to the top andbase section and the wall element of the container is composed of aflexible material. Alternatively the container may comprise a syringearrangement allowing it to be re-filled or emptied. In such a syringearrangement, the container has an arrangement analogous to a syringehaving an element that is moveable to either expel fluid from thecontainer or draw it back in.

In certain embodiments, the container may comprise any shaped containerwith a moving seal allowing variable volume operations.

In certain embodiments the primary container is compressible.

In certain embodiments the primary container comprises a base section, atop section arranged substantially in parallel with the base section anda wall element arranged between the top section and the base section anddefining an internal lumen of the container, in which the wall elementof the container preferably is compressible with respect to the top andbase section and the wall element of the container is composed of aflexible material.

In certain embodiments the container(s) is one of: a reagent container,a bioreactor, a cell culture container, a waste container, a filter, anelectroporator, a purifier, a waste container, a filter, anelectroporator, a purifier, holding container, apheresis/leukopheresis,differentiation chamber, chromatography column, settling chamber, sieve,shaking/mixer a centrifuge and a magnetic bead separator or the like.

In certain embodiments the primary container is a cell culturecontainer.

In certain embodiments control of the cell processing unit is automated.

The term “automated” is used to refer to operation of a componentwithout, or substantially without, user intervention.

In certain embodiments the cell processing unit comprises a controlsystem operable to activate the actuator and/or the drive means. In thisway, a cell processing device loaded into the unit can be selectivelymoved to position a container in line with the actuator and/or theactuator activated to act upon a container in the housing so as to expelits contents.

In certain embodiments the control system is manually or automatic. Morespecifically, the automatic control system may be programmed to operatethe actuator and/or the drive means in a predetermined sequence.

In certain embodiments, the control system comprises a user interface onthe housing.

In certain embodiments, the control system comprises a user interfaceoperably linked to and remote from the housing.

In certain embodiments, the user interface is operable to allow a userto program instructions into the control system.

In certain embodiments the cell processing unit comprises a temperaturecontrol means. In this way, the temperature within the housing can becontrolled and selected.

According to another aspect the disclosure provides a cell processingsystem comprising a cell processing unit according to the disclosure.

According to a yet further aspect of the disclosure there is provided amethod of cell and/or gene therapy manufacture utilizing a cellprocessing unit according to the disclosure.

As will be clear to the person skilled in the art, elements, components,features and advantages of the cell processing unit, cell processingplatform, cell processing device, cell processing container, sterileconnector ends, and the methods of manufacture, usage and componentsthereof may be applied equally to various embodiments described herein.That is, where a feature is described in relation to one embodiment,aspect or example, this is not intended to preclude the inclusion ofsuch a feature in relation to another embodiment, aspect or example, aswill be recognized by those skilled in the art.

BRIEF DESCRIPTION OF THE DRAWINGS

These and other aspects, features and advantages of which embodiments ofthe disclosure are capable of, will be apparent and elucidated from thefollowing description of embodiments and aspects of the disclosure,reference being made to the accompanying drawings, in which:

FIG. 1 illustrates a perspective view of a cell processing unitaccording to an embodiment of the disclosure with a cell processingdevice partially loaded into the device;

FIG. 2 illustrates a side view of a cell processing device according toan embodiment of the disclosure;

FIG. 3 illustrates a cross-sectional view of a part of the cellprocessing device of FIG. 2;

FIGS. 4a and 4b illustrate a perspective view of the valve means of thecell processing platform of the cell processing device of FIG. 2;

FIG. 5 illustrates an isolated side view of one auxiliary container portand auxiliary container of the cell processing device FIG. 2;

FIG. 6 illustrates a perspective view of the mounting bracket, actuatorsand frictional drive mechanism of the cell processing unit of FIG. 1;

FIG. 7 illustrates a top view of the mounting plate and the frictionaldrive mechanism of the partial cell processing unit of FIG. 6;

FIG. 8 illustrates a perspective view of the underside of the cellprocessing device of FIG. 2;

FIG. 9 illustrates a close up view of the cell processing device andsensor arrangement of FIG. 8;

FIG. 10 illustrates a top view of the cell processing device and sensorarrangement of FIG. 8;

FIG. 11 illustrates a Hall Effect Sensor of the cell processing unit anda cell processing platform comprising at least one magnet;

FIG. 12 shows a perspective view from the side of a representation ofone embodiment of a cell processing container comprising a plurality ofsterile connectors according to an embodiment of the disclosure;

FIG. 13 shows a perspective view from the side of a representation ofone embodiment of the cell processing system of the disclosure;

FIG. 14A shows a perspective view from the side of a representation ofone embodiment of the cell processing system of the disclosure, whereauxiliary containers are connected to the cell processing container,leaving an empty auxiliary container port for a further auxiliarycontainer to be connected;

FIG. 14B shows a perspective view from the side of a representation ofone embodiment of the cell processing system of the disclosure, where anauxiliary container has been connected to the empty auxiliary containerport of the cell processing container;

FIGS. 15A, 15B, 15C and 15D show a known sterile connector arrangementformed from two sterile connector ends;

FIGS. 16A, 16B, 16C and 16D show the formation of a sterile connectorfrom two known sterile connector ends;

FIG. 17A shows a perspective view from the side of a representation ofone embodiment of a cell processing container comprising a sterileconnector end embedded therein;

FIG. 17B shows a close view of the sterile connector end of the cellprocessing container of FIG. 15A;

FIGS. 17C, 17 d and 17E a perspective view from the side of arepresentation of an auxiliary container for a cell processing deviceand/or a cell processing system according to the disclosure comprising asterile connector end and being prepared for filling with reagent;

FIG. 18A shows a perspective view from the side of a representation ofone embodiment of an auxiliary container comprising a sterile connectorend embedded in a base section and a screw top cap in a top section;

FIG. 18B shows a perspective view from the side of a representation ofone embodiment of a cell processing container comprising a plurality ofsterile connector ends embedded in a top and a bottom section;

FIG. 18C shows a schematic representation of a number of prefilledauxiliary containers being connected to a cell processing container tocreate a cell processing system according to the disclosure having asterile connector end in an auxiliary container port for receiving afurther auxiliary container containing patient cells;

FIG. 18D shows a schematic representation of a number of prefilledauxiliary containers being connected to a single use wave container tocreate a cell processing system according to the disclosure having asterile connector end in an auxiliary container port for receiving afurther auxiliary container containing patient cells; and

FIG. 18E shows a schematic representation of a number of prefilledauxiliary containers being connected to a CSTR bioreactor to create acell processing system according to the disclosure having a sterileconnector end in an auxiliary container port for receiving a furtherauxiliary container containing patient cells.

DETAILED DESCRIPTION

Specific embodiments of the disclosure will now be described withreference to the accompanying drawings. This disclosure may, however, beembodied in many different forms and should not be construed as limitedto the embodiments set forth herein; rather, these embodiments areprovided so that this disclosure will be thorough and complete, and willfully convey the scope of the disclosure to those skilled in the art.The terminology used in the detailed description of the embodimentsillustrated in the accompanying drawings is not intended to be limitingof the disclosure. In the drawings, like numbers refer to like elements.

The terminology used herein is for the purpose of describing particularaspects of the disclosure only, and is not intended to limit thedisclosure. As used herein, the singular forms “a,” “an” and “the” areintended to include the plural forms as well, unless the context clearlyindicates otherwise.

In the drawings and specification, there have been disclosed exemplaryaspects of the disclosure. However, many variations and modificationscan be made to these aspects without substantially departing from theprinciples of the present disclosure. Thus, the disclosure should beregarded as illustrative rather than restrictive, and not as beinglimited to the particular aspects discussed above. Accordingly, althoughspecific terms are employed, they are used in a generic and descriptivesense only and not for purposes of limitation, for example, definitionof dimensions such as width or breadth or height or length or diameterdepends on how exemplary aspects are depicted, hence, if depicteddifferently, a shown width or diameter in one depiction is a length orthickness in another depiction.

It should be noted that the word “comprising” does not necessarilyexclude the presence of other elements or steps than those listed andthe words “a” or “an” preceding an element do not exclude the presenceof a plurality of such elements. It should further be noted that anyreference signs do not limit the scope of the claims, that the exampleaspects may be implemented at least in part by means of both hardwareand software, and that several “means,” “units” or “devices” may berepresented by the same item of hardware.

Features, integers, characteristics, compounds, chemical moieties orgroups described in conjunction with a particular aspect, embodiment orexample of the disclosure are to be understood to be applicable to anyother aspect, embodiment or example described herein unless incompatibletherewith. All of the features disclosed in this specification(including any accompanying claims, abstract and drawings), and/or allof the steps of any method or process so disclosed, may be combined inany combination, except combinations where at least some of suchfeatures and/or steps are mutually exclusive. The disclosure is notrestricted to the details of any foregoing embodiments. The disclosureextends to any novel one, or any novel combination, of the featuresdisclosed in this specification (including any accompanying claims,abstract and drawings), or to any novel one, or any novel combination,of the steps of any method or process so disclosed.

In the drawings like reference numerals refer to like parts.

Cell Processing Unit

FIG. 1 illustrates a cell processing unit 1 according to the disclosure.The cell processing unit comprises a housing 2 formed of four wallsupstanding from a base wall and a top wall parallel to the base wall andspaced apart from it by the length of the walls. The housing 2 forming achamber 3 with a hinged door 7 in one wall for receiving a cellprocessing device 901 comprising cell processing platform (CPP) 9. Onthe front panel of the cell processing unit 1 is a control panel 5 toenable the user to program and control various features positionedwithin the chamber 3, as well as their interactions with the cellprocessing device 901. Details of these features and the cell processingdevice 901 are set out in more detail below.

The cell processing unit 1 has a housing 2, which defines an enclosedspace, being chamber 3 in which one or more unit operations (i.e.,steps) of cell and/or gene therapy manufacturing process can occur.

An automated cell processing system according to an embodiment of thedisclosure comprises cell processing unit 1 and a cell processing device901 as shown in FIG. 2. The cell processing device 901 comprises a cellprocessing platform 9 and one or more auxiliary containers 11 coupled tothe platform 9. The platform 9 can be manipulated by the cell processingunit 1 to transfer liquids between the auxiliary container 11 (e.g.,feed bellows) located on the top of the cell processing platform 9 andthe primary container 13 (e.g., reactor bellow) located on the bottom ofthe cell processing platform 9. FIG. 1 shows an embodiment in which thecell processing system has cell processing unit 1 and a cell processingdevice 901 with five auxiliary containers 11 fluidly connected to thecell processing platform 9. The cell processing unit 1 rotates theplatform 9 using a friction drive system. The cell processing unit 1comprises a valve solenoid micro-linear actuator (38, FIG. 6), whichwhen activated, opens pinch valves 27 in the cell processing platform 9and presses the auxiliary container 11 using a linear actuator (106,FIG. 6). The cell processing platform 9 comprises a body portioncomprising base plate 15 onto which the primary container 13 (e.g.,reactor bellow) is fitted on the underneath into a primary containerport (FIG. 2, reference numeral 14) and the five auxiliary containers 11(e.g., feed bellows) are fitted on top of the base plate 15 in auxiliarycontainer ports 19. The auxiliary containers 11 (e.g., feed bellows) aremounted on top of the sleeves forming the auxiliary container ports 19,which contain LUER-LOK™ fittings to connect the auxiliary containers 11to the tubing in the auxiliary container ports 19. The tubing beingfluidly connected to the tubing in the base plate 15, through the baseplate 15 and onto the fluid outlet at the primary container port 14.Each auxiliary container port 19 comprises a filling valve 31, whichallows for filling of the auxiliary container 11 fluidly coupled to theport 19. The base plate 15 of the cell processing platform 9 containsnormally closed pinch valves 27 acting on the tubing 29 between theauxiliary containers 11 and the primary container 13. In thisembodiment, the cell processing system comprises a cell processingdevice with five auxiliary containers. However, it should be appreciatedthat in cell processing device may have a different number of auxiliarycontainers according to the present disclosure. It is further envisagedthat the containers may have different volumes according to the presentdisclosure.

The chamber 3 is not sterile, however the containers are completelyclosed when loaded into the cell processing platform. The containers inparallel and/or series in the cell processing platform provide a singleclosed consumable unit (cell processing device) for the entiremanufacturing process. Filling the containers occurs either aseptically(e.g., in a laminar flow hood) or using sterile connections (e.g., tubewelding or sterile connections).

The housing 2 of the cell processing unit 1 allows for easy insertionand removal of the cell processing device 901 through a front openingdoor 7. With the door 7 open, the cell processing device 901 comprisingthe cell processing platform 9 and attached auxiliary containers 11 eachcomprising various cell processing reagents can be placed down and slidinto its final position. The control panel 5 is located on the front ofthe housing 2, meaning that all interactions with the cell processingunit 1 happen from the front. In this way, multiple cell processingunits 1 can be placed close together, side by side or on top of eachother. Having rows of units or stacks of units, respectively,facilitates the capacity for advanced manufacturing and processing. Thedepicted embodiment is shown with five buttons, one for each feedactuation in a test protocol for the system. The door 7 is transparentso that the operations can be visible when demonstrating the function ofthe apparatus. In alternative embodiments an opaque door could beprovided. In this way, the cells can be shielded from UV light duringprocessing.

Cell Processing Unit

FIG. 6 shows a portion 101 of cell processing unit 1 with the housing 2removed for ease of depiction. Inside the housing the portion of thecell processing unit 101 comprises a linear actuator 103 for compressionof the auxiliary container 11 feed bellows, a linear actuator 106 forcompression of the primary container 13 reactor bellow, a frictionaldrive mechanism (107, 109, 111) mounted on mounting plate 104 andoperable to rotate the cell processing platform 9 and a micro linearactuator 38 for opening the pinch valves, which are operable to open andclose the tubing in the platform. The internal structure of theapparatus is machined from aluminum, the linear actuators 106, 103 arealuminum and steel constructions with the lead screws hard coated in TFEdry lubricant.

In addition to the mounting plate 104, the mounting bracket comprises amounting flange (not shown), located above the mounting plate in such away as to retain the cell processing platform by frictional fit betweenthe mounting plate 104 and the mounting flange.

The layout of the actuators 38, 103, 106 allows them to be hidden in therear of the apparatus by a cover (not shown) through which only theplungers 103 a, 106 a protrude to compress the bellows of the auxiliaryand primary containers respectively, helping to give a clean anduncomplicated appearance, and provides an apparatus that is simpler toclean and wipe down. A power supply and the electronics for theactuators and the frictional drive mechanism are mounted on the plate112 below the mounting plate 104. The four risers 114 are adjustable inheight and operable to change the distance between the mounting plate104 and the plate 112 housing the power supply and the electronics. Inthis way, the apparatus can accommodation different sizes of primarycontainers.

The housing 2 contains all of the actuators and electronics necessary tomanipulate the cell processing device. The feed bellow plunger 103 a andreactor plunger 106 a operable to exert a compression force on theauxiliary container and the primary container respectively, attach tolinear rails, each with a maximum force of 100N. The motors driving thelinear rails are bipolar stepper motors. The valve actuator 38 is alinear actuator with a maximum force of 45N.

The frictional drive mechanism (107, 109, 111) comprises a drive wheel107 located on mounting plate 104 and operable to impart rotation on thecell processing device. The drive wheel 107 is a bipolar stepper motor.The actuator stepper motors on the linear rails and the stepper motor inthe frictional drive mechanism are driven by a control system andassociated power supply (not shown).

FIG. 7 shows the elements of the frictional drive mechanism (107, 109,111) mounted to the mounting plate 104 of the mounting bracket. To allowthe cell processing device 901 comprising the cell processing platform 9and the auxiliary containers to be inserted from front only, a drivemethod has been developed where the cell processing platform 9 is heldbetween three friction wheels, one of which being driven 107, the otherdrive mechanism 109 and the third being a hinge wheel 111 within thedoor, which opens to allow insertion of the platform 9 and closes tolock it in place. The cell processing device 901 rotates on low frictionPTFE pads 116 on the mounting plate 104. The spring force of the sprungfriction wheel 109 will be such that there is no slip between the drivewheel 107 and the outer face of the base plate 15 of the platform 9. Thedriven wheel 107 is directly connected to a stepper motor. The baseplate 15 of the cell processing platform 9 is fitted with a series ofmagnets 118 around its circumference so that its position can be read bya Hall Effect sensor 120 mounted on the mounting plate 104. The cellprocessing platform 9 therefore acts like an encoder and gives closedloop position feedback independent of any motor slip.

The Hall Effect sensor 120 mounted to the mounting plate 104 attached tothe housing 2 is operable to detect the magnetic field from the magnets118 on a cell processing platform 9 mounted in the housing 2. The HallEffect sensor 120 is operable to detect the position of the cellprocessing platform 9 relative to the mounting bracket. As best seen inFIG. 8, each auxiliary container port 19 attached to the base plate 15of the cell processing platform 9 has a magnet 118 positioned in thebase plate 15 adjacent the port 19. In this way, the Hall Effect sensor120 will detect a magnet 118 when an auxiliary container port 19 and itsassociated magnet 118 is in line with the sensor. Therefore therespective auxiliary container port 19 is in a known position in thehousing relative to the mounting plate 104.

FIGS. 8, 9, 10 and 11 show the positional sensor array operable todetect the position of the cell processing platform 9 of the cellprocessing device within the cell processing unit 1.

The sensor array comprises Hall Effect sensors 120 and a series ofmagnets 118 on the base plate 15. The sensor array tracks the positionof the cell processing platform 9 using the Hall Effect sensors 120. TheHall Effect sensors 120 produces a voltage in response to magneticfields produced by magnets 118. There are two Hall Effect sensors 120mounted to the mounting plate 104 in the housing 2 and a series ofmagnets 118 embedded in the cell processing platform 9. One of the HallEffect sensors 120 is for tracking rotation of the cell processingplatform 9 relative to the mounting plate 104 and the other Hall Effectsensor 120 is dedicated to tracking a so-called home position of thecell processing platform 9 relative to the mounting plate 104. The homeposition is determined by having one magnet 118 on a different pitchcircle diameter to the other magnets 118 on the cell processing platform9, serving as an index or marker to count full revolutions of the cellprocessing platform 9 in the housing 2. Using the cell processing deviceas an encoder, rather than having an encoder on the motor, means thatthere is a closed loop position feedback on the cell processing deviceitself

To ensure there will be no slip between the drive mechanism and theplatform 9, the friction between the elastomeric driving (friction)wheel 107 and the base plate 15 needs to be greater than the frictionbetween the PTFE pads 116 and the base plate 15. Using the maximum forcethat will be transmitted between the drive wheel 107 and the base plate15 of the platform 9, the normal force required to ensure consistentdrive can be calculated.

Cell Processing Device

The cell processing platform 9, as shown in FIGS. 2-5, comprises a cellprocessing platform having an annular base plate 15 with a number ofauxiliary container ports 19, in this case five, arranged on the uppersurface, and a single primary or reaction container 13 mounted on itsunderside at a primary container port 14. Each auxiliary container port19 is adapted to receive an auxiliary container 11, such as the typesdescribed herein, or in the earlier publication WO2018087558. Each ofthe auxiliary containers 11 in the example has a 45 ml maximum capacitysuch that the total feed capacity of the five auxiliary containers 11 is225 ml. The primary container 13 has a maximum capacity of 150 ml.

As shown in the cross-section of FIG. 3, the auxiliary container 11comprises a top section 21 and a base section 23 with a collapsiblebellows portion 17 located between them to define a storage volume 20.The base section 23 includes fluid outlet 25 through which the contentsof the storage volume 20 can be transferred. With the auxiliarycontainer 11 located into auxiliary container port 19, the outlet 25 isin fluid communication with a connector 26 located therein. In theexample shown, the connector 26 comprises a 4-way stopcock described inmore detail below.

The auxiliary containers 11 are formed of blow molded LDPE while theauxiliary container ports 19 are formed of Nylon. The base plate 15 isformed of machined HDPE and the primary container 13 is formed of blowmolded HDPE bonded to a machined HDPE flange being the primary containerport 14. The base plate 15 is made up of three pieces, which are screwedtogether. The primary container 13 is mounted to the base plate 15 byscrews.

A flexible tubing 29 comprises a first end fitted to connector 26, and asecond end fitted to base plate outlet 33, thereby forming a fluidcommunication conduit between the auxiliary container 11 and the primarycontainer 13. The flexible tubing 29 may comprise any appropriate lengthand cross section. In the example show, the flexible tubing 29 isCOLE-PARMER® Platinum Cured Silicone Tubing with inner diameter (ID) ⅛″and outer diameter (OD) 3/16″. Aptly, the flexible tubing will be madefrom a suitably non-leachable, resilient and biologically inertmaterial, in this case silicone, although other resilient materials maybe used.

Fluid flow through the fluid communication conduit, and hence between anauxiliary container 11 and the primary container 13 is controlled byvalve means 27, located within the base plate 15. In the example shown,the auxiliary container 11 is one of several, each located in acorresponding auxiliary container port 19 on the base plate 15.Accordingly, each auxiliary container 11 is provided with a flexibletubing 29 to the primary container 13, controlled by a separate valvemeans 27. In this way, the transfer of the contents of each storagevolume 20 may be precisely and independently controlled.

One of the valve means 27 is shown in more detail in FIGS. 4a and 4b .The valve means 27 comprises a closure portion 37 slidably engagedwithin a radial channel located in the base plate 15 and defined betweenchannel walls 41 a and 41 b. The closure portion 37 is substantially ahollow rectangular shape with the longer pair of opposing walls arrangedparallel with the channel walls 41 a and 41 b and the shorter pair ofopposing walls arranged at its inner and outer surfaces. An actuatingportion 38 is provided on the outer short wall and a compression portion43 is provided on the inner shorter wall.

The closure portion 37 is the located over a valve wall 39 fixed withinthe channel and spaced away from the channel walls 41 a and 41 b. Theclosure portion 37 can thus be moved between two extreme positions—aclosed position (FIG. 4a ) and an open position (FIG. 4b )—by slidingpast the valve wall 39 within the channel.

The flexible tubing 29 is arranged to extend through the valve means 27such that a section of the tubing 29 sits between the valve wall 39 andthe compression portion 43. In the closed position, the closure portion37 is urged toward the outer perimeter of the base plate 15 by a spring35. The spring 35 is positioned to act on the compression portion 43,urging it against the flexible tubing 29 and pinching it against thevalve wall 39. Thus, in the closed position, the pinched section oftubing blocks the fluid communication conduit and prevents fluid flow.

To unblock the conduit, the closure portion 37 is moved toward the openposition by pressing the actuating portion 38, releasing the compressionportion 43 from the valve wall 39 and allowing the pinched section ofthe flexible tubing to revert to its original shape and permitting fluidflow.

With the cell processing device installed in the cell processing unit,the valve means 27 is actuated by actuator 38 and opened while theauxiliary container 11 is compressed by plunger 103 a. The actuator 38may be configured so that the valve means 27 opens when the auxiliarycontainer 11 is compressed. Alternatively, actuation may occur as aseparate step, for example, when the auxiliary container 11 is receivedinto the auxiliary container port 19. The actuation may occurautomatically in conjunction with the compression of the auxiliarycontainer 11, or may be controlled to happen independently.

In the example shown, the valve actuation is carried out by a linearactuator 38 located at the rear of the housing 2 of the cell processingunit 1, which acts upon the closure portion 37 to move it toward theopen position. Thus, the valve means is normally closed and actuated toopen only when fluid needs to be delivered to the primary container 13.

As shown in FIG. 3, each auxiliary container 11 is attached to a fillingvalve connector 26 in the form of a 4-way stopcock. The connector 26comprises a LUER-LOK™ port for filling via direct access to theauxiliary container 11. This port, which may be used for manuallyinserting fluids into the auxiliary container, does not have its ownvalve means 27 but is capped instead.

Two further capped LUER-LOK™ ports are provided on base plate 15 forsampling/harvesting fluid, or gas exchange. A first port leads to thehead space of the primary container, while a further port is connectedto the base of the primary container 13.

FIG. 5 depicts the filling port 31 and lever 45 mounted on the auxiliarycontainer port 19. The lever 45 is provided in order to fill theauxiliary container 11 without allowing material to flow into the valvemeans 27 or primary container 13. The lever 45 is operatively connectedto a 4-way stopcock, which forms the connector 26 in the exampledescribed above. At the fill position (lever pointing down), the fillingport 31 is opened and flow of material through the port 31 is directedinto the auxiliary container 11. Then, at the feed position (leverpointing up), the filling port 31 is closed and flow is directed fromthe auxiliary container 11 via the flexible tubing 29 and into theprimary container 13.

Sterile Connectors

FIG. 12 shows a cell processing container 200 according to an embodimentof the disclosure. Cell processing container 200 comprises a basesection 202, a top section 203 and a wall element 204 arranged betweenthe top section 203 and the base section 2. The wall element 204 ispreferably composed of a flexible material. The wall element 204 ispreferably compressible with respect to the top section 203 and the basesection 202. The cell processing container 200 may thereby have a“concertina” or “bellows arrangement,” e.g., it may have one or morez-folds in the wall element 204 arrangement.

The cell processing container 200 may comprise 1 sterile connector endand preferably comprises a plurality of connector ends 205. Theconnector ends 205 are preferably sterile. The sterile connector ends205 are preferably located on the top section 203 and/or on the basesection 202 of the cell processing container 200. The cell processingcontainer 200 preferably comprises at least 1, at least 2, at least 3,at least 4, or at least 5 sterile connector ends 205. According to apreferred embodiment, the sterile connector ends 205 are embedded in thecell processing container 200. The sterile connector ends 205 enable aneasy and sterile connection of auxiliary containers 11 to the cellprocessing container 200.

The cell processing container 200 may have any possible shape. In apreferred embodiment the cell processing container 200 has a circular,square, rectangular, elliptical, or triangular cross section.

In a preferred embodiment, when the cell processing container 200 has acircular shape, the sterile connector ends 205 are preferably connectedto the top 203 and/or base 202 section in an essentially circularpattern. The cell processing container 200 also comprises a sterileconnector end 205 in the center of the top 203 and the base 202. Thesterile connector ends 205 are connected to the top 203 and/or base 202section essentially symmetrically having essentially the same distancebetween the different connector ends 205. This enables an easier andpossibly automated process of cell and/or gene therapy manufacturing. Inan alternative embodiment, when the cell culture container 200 has acircular shape, a sterile connector end 205 are connected to the centerof the top section 203 and base section 202.

An embodiment of the disclosure is shown in FIG. 13 and FIGS. 14A-14B,showing a cell processing system according to the disclosure, comprisinga cell processing container 200 as described above together with one ormore auxiliary containers 11 attached to the cell processing container200. The auxiliary containers 11 are preferably connected to the cellprocessing container 200 via sterile connector ends 205. The auxiliarycontainers 11 are preferably connected to the cell processing container200 on the top section 203 and/or the base section 202. The auxiliarycontainers 11 may also be cell processing containers according to thedisclosure comprising an embedded sterile connector end in a baseportion of the auxiliary container 11.

In further embodiments such as the one shown in FIG. 2, the auxiliarycontainers 11 are fluidly coupled to the cell processing container 13through a body portion 15. The body portion forms part of a cellprocessing platform 9. The auxiliary containers 11 each comprise asterile connector end embedded in the base section of the auxiliarycontainer 11. The embedded sterile connector end interconnects andsealingly engages with a corresponding sterile connector end in the bodyportion 15 of the cell processing platform 9. The cell culture container13, being a primary container, is sealingly engaged with the bottom ofthe body portion 15 so as to form a fluid connection between the bodyportion 15 and the cell culture container 13.

The fluid conduit (not shown) between the sterile connector attachingthe auxiliary container 11 to the body portion 15 and the fluid outlet(not shown) of the body portion 15 to which the cell processingcontainer 13 is attached, comprises a pinch valve. The pinch valve isoperable to open and close the fluid conduit in response to a valveactuator such that, as a compression force is applied to the respectiveauxiliary container 11, the contents of the auxiliary container can betransferred by the application of a compression force to the container.In alternative embodiments, the pinch valve may be replaced by apressure-sensitive valve (e.g., a burst valve) such that the valve opensas a compression force is applied to the respective auxiliary container11.

In the embodiment shown in FIGS. 14A and 14B, one or more of feedbellows 11 are pre-attached to the primary cell processing container 200and prefilled with reagent (e.g., liquid) and stored in a refrigerator.The cell processing system shown in FIGS. 14A and 14B may be used forattaching heat labile components, such as viruses or cells, which needto be stored in at −80 degrees Celsius or in liquid nitrogen. Because,it is expensive to store the whole cell processing system at thesetemperatures, the embedded sterile connector end 205 in the feed bellows11 and in the top of the cell processing container 200 serve as a way toadd the heat labile component(s) without use of an aseptic laminar flowhood or sterile tubing welders thus eliminating tube based connectionsand keeping the system compact.

Advanced blow molding techniques can be used to deposit a second (oreven third), external, coating or layer of plastic impermeable to oxygenonto the wall, top and base of the auxiliary container. In this way,shelf life of the container in storage can be extended.

FIGS. 15A to 15D show an exemplary sterile connection between twosterile connector ends 400. The sterile connector ends 400 each have amechanical connection (such as a screw thread) or latch (not shown)arranged in an internal circumferential manner on the sterile connectorend 400. The internal circumferential latches provide the properorientation of sterile connector end 400 relative to the other to ensurethat the opposedly aligned adhesive members 40 attached to the sterileconnector end 400 achieve a sterile fluid connection. In FIG. 15B, twoadhesive members 40 are aligned so that the front second fold adhesivecoating of each adhesive member 40 mirror each other. This alignment isimportant as the rolled member 40 may be withdrawn in only one lineardirection. Once the two front second fold adhesive coating surfaces arein contact, as shown in FIG. 15C, the member pull grip 50 is pulled awayfrom the longitudinal axis of the sterile conduit 190 thereby exposingthe conduit aperture (FIG. 15D). In FIGS. 15C and 15D, the rolled member40 is completely withdrawn to an unfolded configuration and the conduitapertures are aligned to form a sterile corridor.

In FIG. 16A, two opposing sterile connector ends 150 are aligned so thatthe front second fold adhesive coating 80 of each rolled membrane of thesterile connector ends 150 mirror each other. This alignment isimportant as the rolled membrane may be withdrawn in only one lineardirection. Once the two front second fold adhesive coating 80 surfacesare in contact, as shown in FIG. 16B, the entire adhesive surface areascome into contact thereby sealing each opposing sterile connector ends150 together. In FIG. 16C, the membrane pull grip 50 is pulled away fromthe longitudinal axis of the sterile corridor thereby exposing theconduit aperture 60. In FIG. 16D, the rolled member 40 is completelywithdrawn to an unfolded configuration and the conduit apertures 60 arealigned to form a sterile corridor between each sterile connector end150.

FIG. 17A shows a cell processing container 13 having a sterile connectorend 37 embedded in a top section of the container wall. The sterileconnector end 37 forms one half of a sterile connector when the cellprocessing container 13 is fluidly connected to the correspondingsterile connector end in an auxiliary container 11. In alternativeembodiments, the cell processing container 13 is fluidly connected tothe corresponding sterile connector end in a body portion 15 of a cellprocessing platform 9. The sterile connector end in a body portion 15 ofa cell processing platform 9 being part of a primary container port ofthe platform.

FIG. 17B shows an exploded partial view of the sterile connector end 37of FIG. 17A. FIG. 17B shows a male sterile connector end, being half ofsterile connector, in a top wall of cell culture container 13. Thesterile connector end 37 comprises a removable paper cap which, whenengaged with the removable paper cap of a further sterile connector endis removed, exposes the sterile surfaces enclosed by a screw cap engagedwith screw threads of the sterile connector end 37 and creates a fluidconnection through to the cell processing container lumen. Specifically,the removable paper cap is an anti-contamination pull tab, which isinitially folded over the sterile connector end 37 and has an endprotruding therefrom. The pull tab can then be pulled out to expose thesterile surfaces to each other.

FIGS. 17C to 17E depict an auxiliary container 11 being filled withmedia in a sterile process. The process can be manual or automated. InFIG. 17D the sterile connector end 37 is removed and media filled intothe lumen of the auxiliary container 11. The filling of the auxiliarycontainer 11 is performed under sterile conditions. In FIG. 17E, thesterile connector end 37 is replaced and the auxiliary container 11stored at the appropriate temperature until it is needed for assembly ofthe cell processing system. Once filled and ready for use, the auxiliarycontainer 11 is inverted and the sterile connector end 37 mated andconnected with a corresponding sterile connector end on a primarycontainer such as a cell processing container.

In alternative embodiments such as the one depicted in FIG. 18A, theauxiliary container 11 has a screw cap 51 at one end and a sterileconnector end 37 at the other. In this way, the integrity of the sterileconnector end 37 can be maintained during storage of the auxiliarycontainer 11 by inverting the auxiliary container 11 such that the mediasits at the end of the auxiliary container 11 having the screw cap 51and the sterile connector end 37 is free from any liquid contact.

The embedded sterile connector end 37 ensures that the auxiliarycontainer 11 can be readily connected to an auxiliary container port ofa cell processing platform 9 or directly to a cell processing container13 in a cell processing system according to the disclosure.

FIG. 18A shows an auxiliary container 11 having a sterile connector end37 protected by in an end cap 151 in the base section of the auxiliarycontainer 11. The auxiliary container 11 also has a screw cap 51 in thetop section of the container to allow for filling of the lumen of thecontainer with media or the like. The screw cap 51 is compatible withautomated media filling techniques and apparatus.

The sterile connector end 37 facilitates fluid connection between thelumen of the auxiliary container and the contents in it, with a cellprocessing container 13 having a corresponding sterile connector end ina top section of the container 13. In order to access the sterileconnector end 37 in the base section of the auxiliary container 11, theend cap 151 is removed, the sterile connector end 37 can then be matedinto sealing engagement with a corresponding sterile connector end onthe cell processing container 13. In alternative embodiments, thesterile connector end 37 can be mated into sealing engagement with acorresponding sterile connector end on a cell processing platform. Morespecifically, the sterile connector end 37 can be mated into sealingengagement with a corresponding sterile connector end in the auxiliarycontainer port 19 on a cell processing platform 9.

Advanced blow molding techniques can be used to deposit a second (oreven third), external, coating or layer of plastic impermeable to oxygenonto the wall, top and base of the auxiliary container. In this way,shelf life of the container in storage can be extended.

FIG. 18B shows a cell processing container (reactor bellow) 13comprising a plurality of bottom sterile connectors, being embeddedsterile connector ends 139, in the base section of the cell culturecontainer 13. In the depicted embodiments, the cell processing container13 (e.g., reactor bellow) is fitted with a plurality of sterileconnector ends 141 in a top section of the container 13 for connectionof a plurality of auxiliary containers 11. The auxiliary containers 11may contain media and/or cell nutrients required for cell culture.Alternatively, the auxiliary containers may be for sampling or wasteremoval from cell processing container 13. In a sampling arrangement,the cell processing container (e.g., reactor bellow) 13 may be fluidlyconnected via a pinch valve to a removable auxiliary container 11. Thepinch valve is opened and then the auxiliary container 11 is expanded totake the sample from the cell processing container 13. The pinch valveis then closed before detaching the sample auxiliary container 11. Theconnection could be via LUER-LOK™ or similar, which maintains a sterilebarrier once the pinch valve is closed. Thus, samples may be removedfrom the cell processing container 13. The cell processing container 13(e.g., reactor bellow) is fitted with a plurality of sterile connectorends 139 in a base section of the container 13 for connection to aplurality subsequent collection/processing bellows (not shown). Pinchvalves 127 are housed between the sterile connector ends 141 and thecell culture container 13, which pinch valves 127 can be used to switchon/off the flow of feeds from the auxiliary containers 11. Such valveactivation is useful/necessary, for example, if only partial volumes areneeded or feed needs to be added from a single auxiliary container attwo or more time points.

In alternative embodiments, pinch valves can be embedded in the outlettubing from each auxiliary container 11.

In yet further alternative embodiments, the pinch valves can be pressureactuated to open when compression force is applied to the respectiveauxiliary container 11.

FIG. 18C shows the use of prefilled auxiliary containers 311 in a cellprocessing system 300 according to the disclosure. Four auxiliarycontainers 311 are prefilled with wash buffer and are stored at roomtemperature. Four further auxiliary containers 311 are prefilled withgrowth media and are stored at 4 degrees Celsius. Five auxiliarycontainers 311 are prefilled with Lentivirus are stored at −80 degreesCelsius. Four further auxiliary containers 311 are prefilled with mediaincorporating magnetic beads and stored at 4 degrees Celsius. One eachof the prefilled auxiliary containers 311 are connected to the cellprocessing container 313 via sterile connector ends embedded in the baseportion of each auxiliary container 311 and in the top of the cellculture auxiliary container 311. An auxiliary container port 319 remainsempty and ready for receiving a container including patient cells. Itshould be appreciated that in alternative embodiments, the cellprocessing system 300 comprises a different number of prefilledauxiliary containers 311 according to the present disclosure. Forexample, each set of prefilled auxiliary containers 311 may comprise 10sor even 100s of auxiliary containers 311.

The cell processing system including the auxiliary containers 311 andthe cell processing container 313 is now ready for processing in a cellprocessing unit according to the disclosure.

FIGS. 18D and 18E shows the prefilled auxiliary containers 311 housed ona conventional single use wave bioreactor 413 and CSTR bioreactor 513.

The cell processing unit, cell processing platform, cell processingdevice and cell processing container according to the disclosure may beused in any chemical, biological or separation process. Unit processes(e.g., steps) of such processes may be undertaken. The cell processingdevice , in conjunction with the cell processing unit and, optionally,at least one cell processing container of the disclosure may be used incell culture processes (e.g., culturing, manipulating, expanding orstoring cells) or in gene modification processes (e.g., steps includingpurifying, genetically modifying, recovery and wash processes). Othersuitable unit processes that can be performed in the cell processingunit, platform, device and container of the disclosure include but arenot limited to purification (e.g., affinity, size), washing, settling,centrifugation, filtration, chromatography, magnetic bead processes,transduction, electroporation, novel hydrogels, shipping and thawing,expansion of cells in culture, genetic modification andcryopreservation.

A cell processing device and a cell processing container of thedisclosure are each suitable for cell culture and processing of cells,including the use of the container in cell therapy, gene therapy vectorproduction and/or exosome production. A container or device of thedisclosure may be suitably sterilized prior to use (e.g., by gammairradiation or other means). Optionally, the internal surface of thecontainer may be coated with or comprise biologically active agents,which can act on the cells in culture and/or induce differentiation.

The cell processing equipment described herein may be used in cellmanufacturing and/or gene therapy manufacturing processes involving anysuitable cell or gene type. For example, the device of the disclosuremay be used to culture any prokaryotic or eukaryotic cell, suitably ananimal cell, e.g., a mammalian, cell. The cells may be human ornon-human. Examples of sources of suitable non-human cells include,rodents such as mice, rats, and guinea-pigs, as well as ungulate animalsselected from ovine, caprine, porcine, bovine and/or equine species, ornon-human primate species. However, the cells may be bacteria, yeast,fungi or plant cell in origin also.

The cells may be of any type including somatic cells and non-somaticcells. The cells may be stem cells derived from any stage of developmentof the embryo, foetus or adult animal. The cells may be geneticallymodified cells, such as chimeric antigen receptor T-cells (CARTs). Thecells may be from a deposited cell line, such as genetically-modifiedChinese Hamster Ovary (CHO) cells to produce recombinant proteins.

For example, embryonic stem (ES) cells, including cells of non-humanorigin. The cells may be derived from a deposited cell line, such as anES cell line. The ES cells may be derived from means that do notnecessitate the destruction of a human embryo such as parthenogeneticactivation, as described in WO 2003/046141. The cells may be cells of acancer or a hybridoma, which can be caused to proliferate in cultureand/or produce monoclonal antibodies. The cells may also be derived fromthe result of somatic cell nuclear transfer (SCNT) in which the nucleusof a somatic cell is placed into an enucleated oocyte.

The cells may be pluripotent stem cells, for example, primatepluripotent stem (pPS) cells, for example, human embryonic stem (hES)cells. Where the cells are stem cells, the source may be from any tissueof the body, including mesenchymal stem cells (including umbilical cordderived stem cells), neural stem cells or haematopoietic stem cells.Also included are induced pluripotent stem (iPS) cells.

The disclosure therefore provides for the processing of cells within asingle device with multiple unit processes taking place as desiredwithin the cell processing device via delivery/extraction of desiredreagents, waste, cells, or product into or from one or more auxiliarycontainers in fluid communication with the primary container, therebyavoiding the risk of contamination. The system is simpler to use andfurther avoids the complexity of existing approaches. The disclosureprovides for the safer processing of cells with improved reproducibilityand ease of use.

The disclosure also provides for the extraction of cells from a patient(biopsy, such as blood or bone marrow), separation of cells, processingof cells (including cytokine stimulation and/or genetic modifications),solid-liquid separations and loading into a delivery device where thecells can be cultured in the same device throughout the entire process.

In embodiments of the disclosure, cell processing containers forperforming unit operations in cell and/or gene therapy manufacturing canbe assembled in any configuration. In this way, a cell processing systemmay be provided within which a wide variety of processes (bothbiological, chemical and separations) can be undertaken. Similarly, thecell processing system may comprise a cell processing platform of thedisclosure in conjunction with one or more cell processing containers.In this way it is possible to provide a multistage bioreactor operableto perform one or more unit operations in cell and/or gene therapymanufacturing. Because each cell processing container is based on aconcertina arrangement (which can act as a pump) there is no need forpumps and complex sets of tubing/pipes. The system therefore shrinks thespace needed for any given manufacturing process. A cell processingsystem according to the disclosure is particularly well suited forautologous (patient specific) cell and gene therapy where one needs torun a whole manufacturing run for each patient. Using traditionalmanufacturing approaches is not feasible when scaling up to over 5000patients/year given the amount of space needed to run so many parallelmanufacturing runs.

Certain embodiments of the disclosure are described in the followingnumbered clauses.

In certain embodiments, unless mutually incompatible, any one or more ofthe features of one numbered clause may be combined with any one or moreof the features of any other one or more of the numbered clauses. Morespecifically, any one of clauses 1 to 31 may be combined with one ormore of clauses 32 to 47 unless features are mutually incompatible.Further, any one of clauses 1 to 31 may be combined with one or more ofclauses 48 to 76 unless features are mutually incompatible. Yet furtherany one of clauses 32 to 47 may be combined with one or more of clauses48 to 76 unless features are mutually incompatible.

In certain embodiments, unless mutually incompatible, any one or more ofthe following numbered clauses may be combined with any one or more ofthe appended claims.

1. A cell processing unit for cell and gene therapy manufacturecomprising a housing defining an enclosure into which a cell processingplatform can be mounted, a platform mounting bracket within the housingand configured and arranged to receive and retain a cell processingplatform, a drive apparatus configured and arranged to operativelyengage and act upon a cell processing platform so as to move same withrespect to the platform mounting bracket, and an actuator configured andarranged to exert a force on a container mounted into the cellprocessing platform so as to expel a contents from the container.

2. A cell processing unit according to clause 1, wherein the platformmounting bracket comprises a mounting plate.

3. A cell processing unit according to clause 2, wherein the platformmounting bracket comprises a retaining flange spaced apart from themounting plate in order that a cell processing platform can be receivedand retained in position in the housing between the mounting plate andthe retaining flange.

4. A cell processing unit according to clause 2 or clause 3, wherein themounting plate is substantially C-shaped.

5. A cell processing unit according to any one of the preceding clauses,wherein the drive apparatus is a rotational drive apparatus configuredand arranged to operatively engage and act upon a cell processingplatform so as to rotate same with respect to the platform mountingbracket.

6. A cell processing unit according to clause 5, wherein the rotationaldrive apparatus comprises a drive wheel, which is mounted on theplatform mounting bracket and is configured to engage a surface of acell processing platform and to impart rotational movement on it.

7. A cell processing unit according to clause 6, wherein the rotationaldrive apparatus comprises a sprung wheel biased toward the drive wheeland spaced apart from it and mounted on the platform mounting bracket.

8. A cell processing unit according to clause 6 or clause 7, wherein therotational drive apparatus comprises a hinged wheel biased toward thedrive wheel and spaced apart from it and mounted on the platformmounting bracket.

9. A cell processing unit according to clause 8, wherein the hingedwheel is moveable into an open position in which a cell processingplatform can be inserted into and engaged with the cell processingplatform mounting bracket and a closed position in which the hingedwheel is engaged with a surface of the cell processing platform in orderto retain same in the cell processing platform mounting bracket.

10. A cell processing unit according to any one of the precedingclauses, wherein the actuator is a linear actuator.

11. A cell processing unit according to clause 10, wherein the linearactuator comprises a plunger operatively coupled to a drive motor,wherein the plunger is configured to engage a container in the cellprocessing platform and to exert a compression force on the container.

12. A cell processing unit according to any one of the precedingclauses, comprising a primary actuator configured and arranged to exerta force on a primary container mounted to the cell processing platformso as to expel a fluid from the container.

13. A cell processing unit according to clause 12, wherein the primaryactuator is a linear actuator.

14. A cell processing unit according to clause 13, wherein the primaryactuator comprises a plunger operatively coupled to a drive motor,wherein the plunger is configured to engage a primary container mountedto the cell processing platform and to exert a compression force on theprimary container.

15. A cell processing unit according to any one of the precedingclauses, comprising a valve actuator operable to act upon a pinch valvein the cell processing platform so as to open and close same as force isapplied to the container.

16. A cell processing unit according to clause 15, wherein the valveactuator is a linear actuator.

17. A cell processing unit according to clause 16, wherein the valveactuator comprises a solenoid valve.

18. A cell processing unit according to any one of the precedingclauses, comprising a location detecting sensor operable to detect theposition of the cell processing platform relative to the platformmounting bracket.

19. A cell processing unit according to clause 18, wherein the locationdetecting sensor is operable to detect the rotational position of thecell processing platform relative to the platform mounting bracket.

20. A cell processing unit according to clause 18 or clause 19, whereinthe location detecting sensor comprises a Hall Effect sensor.

22. A cell processing unit according to any one of clauses 18 to 20,comprising a home location detecting sensor operable to detect a homeposition of the cell processing platform relative to the platformmounting bracket.

22. A cell processing unit according to clause 21, wherein the homelocation detecting sensor is operable to detect a single rotationalposition of the cell processing platform relative to the platformmounting bracket.

23. A cell processing unit according to clause 21 or clause 22, whereinthe home location detecting sensor comprises a Hall Effect sensor.

24. A cell processing unit according to clause 23, wherein the voltagedetected by the Hall Effect sensor is greater at the home position ofthe cell processing platform relative to the platform mounting bracketthan at any other position during the rotation of the cell processingplatform relative to the platform mounting bracket.

25. A cell processing unit according to any one of the precedingclauses, wherein the container is compressible.

26. A cell processing unit according to any one of the precedingclauses, wherein the container comprises a base section, a top sectionarranged substantially in parallel with the base section and a wallelement arranged between the top section and the base section anddefining an internal lumen of the container, in which the wall elementof the container preferably is compressible with respect to the top andbase section and the wall element of the container is composed of aflexible material.

27. A cell processing unit according to any one of clauses 12 to 26,wherein the primary container is compressible.

28. A cell processing unit according to any one of clauses 12 to 27,wherein the primary container comprises a base section, a top sectionarranged substantially in parallel with the base section and a wallelement arranged between the top section and the base section anddefining an internal lumen of the container, in which the wall elementof the container preferably is compressible with respect to the top andbase section and the wall element of the container is composed of aflexible material.

29. A cell processing unit according to any one of the preceding clauseswhere the container(s) is one of: a reagent container, a bioreactor, acell culture container, a waste container, a filter, an electroporator,a purifier, holding container, apheresis/leukopheresis, differentiationchamber, chromatography column, settling chamber, sieve, shaking/mixer ,a centrifuge and a magnetic bead separator or the like.

30. A cell processing unit according to any one of clauses 12 to 29,wherein the primary container is a cell processing container.

31. A cell processing unit according to any one of the precedingclauses, wherein control of the device is automated.

32. A cell processing container for use in one or more unit operationsin cell and/or gene therapy manufacture, the container having a basesection, a top section arranged substantially in parallel with the basesection and a wall element arranged between the top section and the basesection and defining an internal lumen of the container, in which thewall element of the cell processing container preferably is compressiblewith respect to the top and base section and the wall element of thecell processing container is composed of a flexible material, whereinthe cell processing container comprises at least one sterile connectorend configured to operatively couple with a further sterile connectorend so as to form a sterile connector between the cell processingcontainer and a further component to which the cell processing containeris to be fluidly connected.

33. A cell processing container according to clause 32, wherein the atleast one sterile connector end is a genderless sterile connector endconfigured to operatively couple with a further genderless sterileconnector end.

34. A cell processing container according to clause 32, wherein the atleast one sterile connector end is a male sterile connector endconfigured to operatively couple with a female sterile connector end.

35. A cell processing container according to clause 32, wherein the atleast one sterile connector end is a female sterile connector endconfigured to operatively couple with a male sterile connector end.

36. A cell processing container according to any one of clauses 32 to35, comprising a plurality of sterile connector ends each configured tooperatively couple with a separate further sterile connector end to forma plurality of sterile connectors between the cell processing containerand at least one further component to which the cell processingcontainer is to be fluidly connected.

37. A cell processing container according to any one of clauses 32 to36, wherein the sterile connector ends are embedded in the cellprocessing container.

38. A cell processing container according to any one of clauses 32 to37, wherein the sterile connector end is operatively coupled to a pinchvalve embedded in the cell processing container.

39. A cell processing container according to any one of clauses 32 to38, wherein the cell processing container has a circular, square,rectangular, elliptical, or triangular cross section.

40. A cell processing container according to clause 39, wherein, whenthe cell processing container has a circular shape, the sterileconnector end(s) is/are connected to the top and/or base section of thecell processing container in an essentially circular pattern.

41. A cell processing system, comprising a cell processing containeraccording to any one of clauses 32-40, further comprising one or moreauxiliary containers detachably connected to the cell processingcontainer.

42. A cell processing system according to clause 41, wherein one or moreof the auxiliary containers comprises the further sterile connector endand is connected to the cell processing container via the furthersterile connector end.

43. A cell processing system according to clause 41 or clause 42 whereinone or more of the auxiliary containers is located on the top section ofthe cell processing container.

44. A cell processing system according to clause 41 or clause 42,wherein one or more of the auxiliary containers is located at or nearthe base section of the cell processing container.

45. A cell processing system according to any one of clauses 41 to 44,wherein the one or more auxiliary containers have a base section, a topsection arranged substantially in parallel with the base section and awall element arranged between the top section and the base section anddefining an internal lumen of the container, in which the wall elementof the auxiliary container preferably is compressible with respect tothe top and base section and the wall element of the auxiliary containeris composed of a flexible material.

46. A multi-step method of performing one or more unit operations incell and/or gene therapy manufacture using a cell processing systemaccording to clauses 41-45.

47. The method according to clause 46, comprising introducing a cellpopulation of interest into the cell processing container andsequentially adding one or more reagents from one or more auxiliarycontainers into the cell processing container in order to effect thedesired one or more unit operations in cell and/or gene therapymanufacture.

48. A cell processing platform for use in one or more unit operations incell and/or gene therapy manufacture, the platform comprising a bodyportion comprising at least one fluid inlet fluidly connected to a fluidoutlet, and an auxiliary container port fluidly coupled to the at leastone fluid inlet of the body portion, wherein the auxiliary containerport is configured and arranged to receive and sealingly engage with anauxiliary container and to fluidly connect the auxiliary container lumenwith the at least one fluid inlet of the body portion, and a primarycontainer port configured and arranged to sealingly engage with aprimary container and to fluidly connect the primary container lumenwith the fluid outlet of the body portion.

49. A cell processing platform according to clause 48, wherein theauxiliary container port comprises a container receiving sleeveconnected to the body portion and being configured to surround at leasta portion of the auxiliary container, which portion comprises the fluidoutlet of the container.

50. A cell processing platform according to clause 48 or clause 49,wherein the auxiliary container port comprises a mating elementconfigured to fluidly connect to a corresponding mating element on anauxiliary container.

51. A cell processing platform according to clause 50, wherein themating element is at least one of: a sterile connector end or aLUER-LOK™.

52. A cell processing platform according to any one of clauses 48 to 51,wherein the primary container port comprises a mating element configuredto fluidly connect to a corresponding mating element on a primarycontainer.

53. A cell processing platform according to clause 52, wherein themating element comprises at least one of: a sterile connector end or aLUER-LOK™

54. A cell processing platform according to any one of clauses 48 to 53,wherein the auxiliary container port comprises a sterile connector endat the fluid inlet and/or the fluid outlet of the auxiliary containerport, each sterile connector end configured to engage with a furthersterile connector end on a container and/or on the body portion,respectively.

55. A cell processing platform according to any one of clauses 48 to 54,wherein the fluid outlet of the body portion comprises a sterileconnector end configured to engage with a further sterile connector endon a primary container attachable to the body portion.

56. A cell processing platform according to according to any one ofclauses 48 to 55, wherein the body portion is substantially hollow.

57. A cell processing platform according to according to any one ofclauses 48 to 56, wherein the at least one fluid inlet and the fluidoutlet of the body portion are fluidly coupled to one another by a fluidconduit.

58. A cell processing platform according to clause 57, wherein the fluidconduit comprises a valve operable to open and close the fluid conduit.

59. A cell processing platform according to clause 58, wherein the valveis one of: a pinch valve, a pressure-sensitive valve, a clamp valve, amembrane valve, a rupture disc, a venous valve and an aperture valve.

60. A cell processing platform according to any one of clauses 48 to 59,wherein the auxiliary container port comprises a container filling port.

61. A cell processing platform according to clause 60, wherein thecontainer filling port is fluidly connected to a fluid inlet of theauxiliary container port.

62. A cell processing platform according to clause 60 or clause 61,wherein the container filling port comprises a valve operatively coupledto the fluid inlet and a fluid outlet of the auxiliary container portand operable to control fluid flow direction through the auxiliarycontainer port.

63. A cell processing platform according to any one of clauses 60 to 62,wherein the container filling port comprises a valve operable, in anopen position, to allow fluid to flow to the fluid inlet of theauxiliary container port and not to the fluid outlet of the auxiliarycontainer port and, in a closed position, to close the container fillingport and to allow fluid to flow from the fluid inlet of the auxiliarycontainer port to the fluid outlet of the auxiliary container port.

64. A cell processing platform according to any one of clauses 48 to 63,comprising a plurality of auxiliary container ports each configured andarranged to receive and sealingly engage with an auxiliary container andto fluidly connect the container lumen with a fluid inlet of the bodyportion.

65. A cell processing platform according to clause 64, wherein eachauxiliary container port is coupled to a separate fluid inlet of thebody portion.

66. A cell processing platform according to clause 65, wherein eachseparate fluid inlet of the body portion is fluidly connected to a fluidoutlet of the body portion.

67. A cell processing platform according to any one of clauses 48 to 66,comprising at least one positional tracking device operable to indicatea set location on the platform.

68. A cell processing platform according to clause 67, wherein thepositional tracking device is at least one of: a magnet, an RFID sensor,a light sensor or a cog operable to engage a further cog.

69. A cell processing platform according to clause 67 or clause 68,comprising a plurality of positional tracking devices.

70. A cell processing platform according to any one of clauses 67 to 69,wherein the at least one positional tracking device is located relativeto the auxiliary container port such that the location of the positionaltracking device is related to the position of the auxiliary containerport.

71. A cell processing platform according to any one of clauses 67 to 70,wherein the at least one positional tracking device is located on thebody portion relative to the auxiliary container port.

72. A cell processing platform according to any one of clauses 48 to 71,comprising a sampling port in the body portion.

73. A cell processing platform according to any one of clauses 48 to 72,comprising a gas transfer port in the body portion.

74. A cell processing platform according to any one of clauses 48 to 73,wherein the auxiliary container port is configured to receive acontainer having a base section, a top section arranged substantially inparallel with the base section and a wall element arranged between thetop section and the base section and defining an internal lumen of thecontainer, in which the wall element of the container preferably iscompressible with respect to the top and base section and the wallelement of the container is composed of a flexible material.

75. A cell processing platform according to any one of clauses 48 to 74,wherein the primary container port is configured to receive a primarycontainer having a base section, a top section arranged substantially inparallel with the base section and a wall element arranged between thetop section and the base section and defining an internal lumen of thecontainer, in which the wall element of the container preferably iscompressible with respect to the top and base section and the wallelement of the container is composed of a flexible material.

76. A cell processing platform according to clause 75, wherein theprimary container further comprises an attachment flange mounted to thetop section of the primary container and being configured to sealinglyengage and detachably mount to the primary container port.

1. A cell processing device for use in performing one or more unitprocesses in one or more of cell or gene therapy manufacturing,comprising: a cell processing platform fluidly coupled to at least oneauxiliary container and to at least one primary container, the cellprocessing platform comprising a body portion comprising at least onefluid inlet fluidly connected to a fluid outlet; and an auxiliarycontainer port fluidly coupled to the at least one fluid inlet of thebody portion, wherein the at least one auxiliary container is receivedin sealing engagement with the auxiliary container port to fluidlyconnect the auxiliary container lumen with the at least one fluid inletof the body portion, and a primary container is received in sealinglyengagement with the primary container port to fluidly connect theprimary container lumen with the fluid outlet of the body portion. 2.The cell processing device according to claim 1, wherein the auxiliarycontainer port comprises a container receiving sleeve connected to thebody portion and being configured to surround at least a portion of theauxiliary container, which portion comprises the fluid outlet of thecontainer.
 3. The cell processing device according to claim 1, whereinthe cell processing platform comprises a plurality of auxiliarycontainer ports and wherein each one of a plurality of auxiliarycontainers are received in sealing engagement with one of the pluralityof auxiliary container ports to fluidly couple the lumen of eachauxiliary container is fluidly coupled with a fluid inlet of the bodyportion.
 4. The cell processing device according to claim 3, whereineach auxiliary container port is coupled to a separate fluid inlet ofthe body portion.
 5. The cell processing device according to claim 4,wherein each separate fluid inlet of the body portion is fluidlyconnected to a fluid outlet of the body portion. 6.-12. (canceled) 13.(canceled)
 14. (canceled)
 15. (canceled)
 16. (canceled)
 17. The cellprocessing device according to claim 1, wherein the auxiliary containerport comprises a sterile connector end at one or more of the fluid inletor the fluid outlet of the auxiliary container port, each sterileconnector end configured to engage with a further sterile connector endon a container or on the body portion, respectively.
 18. The cellprocessing device according to claim 1, wherein the fluid outlet of thebody portion comprises a sterile connector end configured to engage witha further sterile connector end on the primary container attachable tothe body portion.
 19. The cell processing device according to claim 1,comprising at least one positional tracking device operable to indicatea set location on the platform.
 20. The cell processing device accordingto claim 19, wherein the positional tracking device is one or more of: amagnet, an RFID sensor, a light sensor or a cog operable to engage afurther cog.
 21. (canceled)
 22. (canceled)
 23. A cell processing deviceaccording to claim 19, wherein the at least one positional trackingdevice is located on the body portion relative to the auxiliarycontainer port.
 24. (canceled)
 25. The cell processing device accordingto claim 1, comprising a sampling port in the body portion.
 26. The cellprocessing device according to claim 1, comprising a gas transfer portin the body portion.
 27. The cell processing device according to claim1, wherein the auxiliary container port is configured to receive anauxiliary container having a base section, a top section arrangedsubstantially in parallel with the base section and a wall elementarranged between the top section and the base section and defining aninternal lumen of the container, in which the wall element of thecontainer preferably is compressible with respect to the top and basesection and the wall element of the container is composed of a flexiblematerial.
 28. The cell processing device according to claim 1, whereinthe primary container port is configured to receive a primary containerhaving a base section, a top section arranged substantially in parallelwith the base section and a wall element arranged between the topsection and the base section and defining an internal lumen of thecontainer, in which the wall element of the container preferably iscompressible with respect to the top and base section and the wallelement of the container is composed of a flexible material.
 29. Thecell processing device according to claim 28, wherein the primarycontainer further comprises an attachment flange mounted to the topsection of the primary container and being configured to sealinglyengage and mount to the primary container port.
 30. The cell processingdevice according to claim 1, wherein, the at least one auxiliarycontainer is compressible.
 31. The cell processing device according toclaim 1, wherein the at least one auxiliary container is one of: asyringe or any shaped container with a moving seal allowing variablevolume operations.
 32. The cell processing device according to claim 1,wherein the at least one auxiliary container is a bag retained in aframe and moveable with respect to the frame.
 33. The cell processingdevice according to claim 1, comprising one or more auxiliary containersdetachably connected to an auxiliary container port of the cellprocessing platform.
 34. The cell processing device according to claim33, wherein one or more of the auxiliary containers are connected to arespective auxiliary container port with a sterile connector. 35.(canceled)
 36. (canceled)
 37. (canceled)
 38. (canceled)
 39. (canceled)40. (canceled)